Biocompatible hydrogel treatments for retinal detachment

ABSTRACT

Provided herein are in vivo gelling ophthalmic pre-formulations forming a biocompatible retinal patch comprising at least one nucleophilic compound or monomer unit, at least one electrophilic compound or monomer unit, and optionally a therapeutic agent and/or viscosity enhancer. In some embodiments, the retinal patch at least partially adheres to the site of a retinal tear. Also provided herein are methods of treating retinal detachment by delivering an in vivo gelling ophthalmic pre-formulation to the site of a retinal tear in human eye, wherein the in vivo gelling ophthalmic pre-formulation forms a retinal patch.

CROSS-REFERENCE

This application is a continuation of U.S. patent application Ser. No.14/273,408, filed May 8, 2014, which was filed pursuant to 35 U.S.C.§111(a) as a continuation of PCT International Application No.PCT/US2013/040619, filed May 10, 2013, which claims the benefit of U.S.Provisional Application No. 61/646,227, filed May 11, 2012, U.S.Provisional Application No. 61/669,577, filed Jul. 9, 2012, and U.S.Provisional Application No. 61/785,358, filed Mar. 14, 2013, which areincorporated herein by reference in their entireties.

BACKGROUND OF THE INVENTION

Retinal detachment is a disorder of the eye in which the retina peelsaway from its underlying layer of support tissue. When the retinabecomes detached, bleeding from area blood vessels may cloud the insideof the eye, which is normally filled with vitreous fluid. Central visionbecomes severely affected if the macula, the part of the retinaresponsible for fine vision, becomes detached. The most common types ofretinal detachments are often due to a tear or hole in the retina. Eyefluids may leak through this opening. This causes the retina to separatefrom the underlying tissues, much like a bubble under wallpaper. This ismost often caused by a condition called posterior vitreous detachment.Another type of retinal detachment is called tractional detachment. Thisis seen in people who have uncontrolled diabetes, previous retinalsurgery, or have chronic inflammation. If not treated in time, itresults in blindness. Initial detachment may be localized, but withoutrapid treatment the entire retina may detach, leading to vision loss andblindness.

Most people with a retinal detachment will need surgery. Surgery may bedone immediately or after a short period of time.

SUMMARY OF THE INVENTION

Provided herein is a in vivo gelling ophthalmic pre-formulation,comprising: (a) at least one first compound comprising more than onenucleophilic group; (b) at least one second compound comprising morethan one electrophilic group; (c) an aqueous buffer in the pH range ofabout 6.0 to about 8.5; and (d) a viscosity enhancer; wherein the invivo gelling ophthalmic formulation at least in part polymerizes and/orgels at a target site of an eye to form a biocompatible retinal patch.In some embodiments, the target site is a retinal tear. In certainembodiments, the biocompatible retinal patch at least partially adheresto the target site. In some embodiments, the in vivo gelling ophthalmicpre-formulation further comprises a therapeutic agent. In someembodiments, the viscosity enhancer is selected fromhydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose,polyvinyl alcohol, or polyvinylpyrrolidone.

In certain embodiments of the in vivo gelling ophthalmicpre-formulation, the nucleophilic group is a thiol or amino group. Insome embodiments, the first compound is a glycol, trimethylolpropane,pentaerythritol, hexaglycerol, or tripentaerythritol derivative. Incertain embodiments, the first compound further comprises one or morepolyethylene glycol sections. In some embodiments, the first compound isa pentaerythritol or hexaglycerol derivative. In certain embodiments,the first compound is selected from the group consisting of ethoxylatedpentaerythritol ethylamine ether, ethoxylated pentaerythritolpropylamine ether, ethoxylated pentaerythritol amino acetate,ethoxylated hexaglycerol ethylamine ether, ethoxylated hexaglycerolpropylamine ether, and ethoxylated hexaglycerol amino acetate. Incertain embodiments, the first compound is selected from the groupconsisting of trimethylolpropane trimercaptoacetate, trimethylolpropanetri-3-mercaptopropionate, pentaerythritol tetramercaptoacetate,pentaerythritol tetra-3-mercaptopropionate, ethoxylatedtrimethylolpropane trimercaptoacetate, ethoxylated trimethylolpropanetri-3-mercaptopropionate, ethoxylated pentaerythritoltetramercaptoacetate, and ethoxylated trimethylolpropanetri-3-mercaptopropionate. In some embodiments, the molecular weight ofthe first compound is between about 100 and 100000. In certainembodiments, the first compound is water soluble. In some embodiments,the electrophilic group is an epoxide, N-succinimidyl succinate,N-succinimidyl glutarate, N-succinimidyl succinamide or N-succinimidylglutaramide.

In certain embodiments of the in vivo gelling ophthalmicpre-formulation, the second compound is a trimethylolpropane, glycerol,diglycerol, pentaerythritol, sorbitol, hexaglycerol, tripentaerythritol,or polyglycerol derivative. In some embodiments, the second compoundfurther comprises one or more polyethylene glycol sections. In certainembodiments, the second compound is a trimethylolpropane,pentaerythritol, or hexaglycerol derivative. In some embodiments, thesecond compound is selected from the group consisting of ethoxylatedpentaerythritol succinimidyl succinate, ethoxylated pentaerythritolsuccinimidyl glutarate, ethoxylated pentaerythritol succinimidylglutaramide, ethoxylated hexaglycerol succinimidyl succinate,ethoxylated hexaglycerol succinimidyl glutarate, and ethoxylatedhexaglycerol succinimidyl glutaramide. In certain embodiments, thesecond compound is selected from the group consisting of sorbitolpolyglycidyl ether, polyglycerol polyglycidyl ether, diglycerolpolyglycidyl ether, glycerol polyglycidyl ether, and trimethylolpropanepolyglycidyl ether. In some embodiments, the molecular weight of thesecond compound is between about 100 and 100000. In certain embodiments,the second compound is water soluble.

In some embodiments of the in vivo gelling ophthalmic pre-formulation,the gelling time of the biocompatible retinal patch is controlled by thepH of the aqueous buffer, the type of the buffer, the concentration ofthe buffer, the concentration of the first compound and/or the secondcompound in the buffer, or the nature of the electrophilic groups. Incertain embodiments, the gelling time is between about 20 seconds and 10minutes. In some embodiments, the pH of the aqueous buffer is from about8. In certain embodiments, the in vivo gelling ophthalmicpre-formulation gels at a predetermined time to form the biocompatibleretinal patch. In some embodiments, the biocompatible retinal patch is abioabsorbable polymer. In certain embodiments, the biocompatible retinalpatch is bioabsorbed within about 1 to 70 days. In some embodiments, thebiocompatible retinal patch is substantially non-bioabsorbable.

In certain embodiments of the in vivo gelling ophthalmicpre-formulation, the therapeutic agent is released from thebiocompatible retinal patch through diffusion, osmosis, degradation ofthe biocompatible retinal patch, or any combination thereof. In someembodiments, the therapeutic agent is initially released from thebiocompatible retinal patch through diffusion and later released throughdegradation of the biocompatible retinal patch. In certain embodiments,the therapeutic agent is substantially released from the biocompatibleretinal patch within 180 days. In some embodiments, the therapeuticagent is substantially released from the biocompatible retinal patchwithin 14 days. In certain embodiments, the therapeutic agent issubstantially released from the biocompatible retinal patch within 24hours. In some embodiments, the therapeutic agent is substantiallyreleased from the biocompatible retinal patch within one hour. Incertain embodiments, the first compound and the second compound do notreact with the therapeutic agent during formation of the biocompatibleretinal patch. In some embodiments, the biocompatible retinal patchinteracts with the therapeutic agent, and wherein more than 10% of thetherapeutic agent is released through degradation of the biocompatibleretinal patch. In certain embodiments, more than 30% of the therapeuticagent is released through degradation of the biocompatible retinalpatch. In some embodiments, the biocompatible retinal patch interactswith the therapeutic agent by forming covalent bonds between thebiocompatible retinal patch and the therapeutic agent. In certainembodiments, the biocompatible retinal patch interacts with thetherapeutic agent by forming a non-covalent bond between thebiocompatible retinal patch and the therapeutic agent. In someembodiments, the therapeutic agent is released while the biocompatibleretinal patch degrades. In certain embodiments, the release of thetherapeutic agent is essentially inhibited until a time that thebiocompatible retinal patch starts to degrade. In some embodiments, thetime the biocompatible retinal patch starts to degrade is longer thehigher a degree of cross-linking of the biocompatible retinal patch. Incertain embodiments, the time the biocompatible retinal patch starts todegrade is shorter the higher a concentration of ester groups in thefirst or second compound.

Also provided herein is a biocompatible retinal patch made by mixing:(a) at least one first compound comprising more than one nucleophilicgroup; (b) at least one second compound comprising more than oneelectrophilic group; (c) an aqueous buffer in the pH range of about 6.0to about 8.5; (d) a viscosity enhancer; and (e) optionally a therapeuticagent; wherein the mixing is performed outside external to a human eye,and the biocompatible retinal patch gels at least in part at a targetsite inside the human eye.

In some embodiments of the biocompatible retinal patch, the viscosityenhancer is selected from hydroxyethylcellulose,hydroxypropylmethylcellulose, methylcellulose, polyvinyl alcohol, orpolyvinylpyrrolidone.

In certain embodiments of the biocompatible retinal patch, the targetsite is a retinal tear. In some embodiments, the biocompatible retinalpatch at least partially adheres to the target site.

In certain embodiments of the biocompatible retinal patch, thenucleophilic group is a thiol or amino group. In some embodiments, thefirst compound is a glycol, trimethylolpropane, pentaerythritol,hexaglycerol, or tripentaerythritol derivative. In certain embodiments,the first compound further comprises one or more polyethylene glycolsections. In some embodiments, the first compound is selected from thegroup consisting of ethoxylated pentaerythritol ethylamine ether,ethoxylated pentaerythritol propylamine ether, ethoxylatedpentaerythritol amino acetate, ethoxylated hexaglycerol ethylamineether, ethoxylated hexaglycerol propylamine ether, ethoxylatedtrimethylolpropane tri-3-mercaptopropionate, ethoxylated hexaglycerolamino acetate.

In some embodiments of the biocompatible retinal patch, theelectrophilic group is an epoxide, N-succinimidyl succinate,N-succinimidyl glutarate, N-succinimidyl succinamide, or N-succinimidylglutaramide. In certain embodiments, the second compound is atrimethylolpropane, glycerol, diglycerol, pentaerythritol, sorbitol,hexaglycerol, tripentaerythritol, or polyglycerol derivative. In someembodiments, the second compound further comprises one or morepolyethylene glycol sections. In certain embodiments, the secondcompound is selected from the group consisting of ethoxylatedpentaerythritol succinimidyl succinate, ethoxylated pentaerythritolsuccinimidyl glutarate, ethoxylated pentaerythritol succinimidylglutaramide, ethoxylated hexaglycerol succinimidyl succinate,ethoxylated hexaglycerol succinimidyl glutarate, ethoxylatedhexaglycerol succinimidyl glutaramide, and sorbitol polyglycidyl ether.

In certain embodiments of the biocompatible retinal patch, the molecularweight of the first compound and the second compound is between about100 and 100000. In some embodiments, the first compound is waterssoluble. In certain embodiments, the second compound is water soluble.

In some embodiments of the biocompatible retinal patch, the gelling timeof the biocompatible retinal patch is controlled by the pH of theaqueous buffer, the type of the buffer, the concentration of the buffer,the concentration of the first compound and/or the second compound inthe buffer, or the nature of the electrophilic groups. In certainembodiments, the gelling time is between about 20 seconds and 10minutes. In some embodiments, the biocompatible retinal patch gels at apredetermined time.

In certain embodiments of the biocompatible retinal patch, thebiocompatible retinal patch is a bioabsorbable polymer. In someembodiments, the biocompatible retinal patch is bioabsorbed within about1 to 70 days. In certain embodiments, the biocompatible retinal patch issubstantially non-bioabsorbable.

In some embodiments of the biocompatible retinal patch, thebiocompatible retinal patch further comprises a radiopaque material or apharmaceutically acceptable dye.

In certain embodiments of the biocompatible retinal patch, thetherapeutic agent is released from the biocompatible retinal patchthrough diffusion, osmosis, degradation of the biocompatible retinalpatch, or any combination thereof. In some embodiments, the therapeuticagent is initially released from the biocompatible retinal patch throughdiffusion and later released through degradation of the biocompatibleretinal patch. In certain embodiments, the therapeutic agent issubstantially released from the biocompatible retinal patch within 180days. In some embodiments, the therapeutic agent is substantiallyreleased from the biocompatible retinal patch within 24 hours.

In certain embodiments of the biocompatible retinal patch, the firstcompound and the second compound do not react with the therapeutic agentduring formation of the biocompatible retinal patch. In someembodiments, the biocompatible retinal patch interacts with thetherapeutic agent, and wherein more than 10% of the therapeutic agent isreleased through degradation of the biocompatible retinal patch. Incertain embodiments, the release of the therapeutic agent is determinedby the composition of the biocompatible retinal patch. In someembodiments, the therapeutic agent is released while the biocompatibleretinal patch degrades. In certain embodiments, the release of thetherapeutic agent is essentially inhibited until a time that thebiocompatible retinal patch starts to degrade. In some embodiments, atleast a portion of the therapeutic agent is released before the timethat the biocompatible retinal patch starts to degrade. In certainembodiments, the time the biocompatible retinal patch starts to degradeis longer the higher a degree of cross-linking of the biocompatibleretinal patch. In some embodiments, the time the biocompatible retinalpatch starts to degrade is shorter the higher a concentration of estergroups in the first or second compound.

Further provided here in a in vivo polymerized biocompatible retinalpatch comprising: (a) at least one first monomeric unit bound through atleast one amide, thioester, or thioether linkage to at least one secondmonomeric unit; (b) at least one second monomeric unit bound to at leastone first monomeric unit; (c) a viscosity enhancer; and (d) optionally atherapeutic agent; wherein the in vivo polymerized biocompatible retinalpatch is polymerized at least in part at a retinal tear in a human eye.In some embodiments, the in vivo polymerized biocompatible patch atleast partially adheres to the retina of the eye. In certainembodiments, the viscosity enhancer is selected fromhydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose,polyvinyl alcohol, or polyvinylpyrrolidone.

In some embodiments of the in vivo polymerized biocompatible retinalpatch, the first monomeric unit is a glycol, trimethylolpropane,pentaerythritol, hexaglycerol, or tripentaerythritol derivative. Incertain embodiments, the first monomeric unit further comprises one ormore polyethylene glycol sections. In some embodiments, the secondmonomeric unit is a trimethylolpropane, glycerol, diglycerol,pentaerythritol, sorbitol, hexaglycerol, tripentaerythritol, orpolyglycerol derivative. In certain embodiments, the second monomericunit comprises one or more polyethylene glycol sections. In someembodiments, the molecular weight of the first monomeric unit and thesecond monomeric unit is between about 100 and 100000.

In certain embodiments of the in vivo polymerized biocompatible retinalpatch, the in vivo polymerized biocompatible retinal patch is abioabsorbable polymer. In some embodiments, the in vivo polymerizedbiocompatible retinal patch is bioabsorbed within about 1 to 70 days. Incertain embodiments, the in vivo polymerized biocompatible retinal patchis substantially non-bioabsorbable.

In some embodiments of the in vivo polymerized biocompatible retinalpatch, the in vivo polymerized biocompatible retinal patch furthercomprises a radiopaque material or a pharmaceutically acceptable dye.

In certain embodiments of the in vivo polymerized biocompatible retinalpatch, the therapeutic agent is released from the in vivo polymerizedbiocompatible retinal patch through diffusion, osmosis, degradation ofthe in vivo polymerized biocompatible retinal patch, or any combinationthereof. In certain embodiments, the therapeutic agent is initiallyreleased from the in vivo polymerized biocompatible retinal patchthrough diffusion and later released through degradation of the in vivopolymerized biocompatible retinal patch. In some embodiments, thetherapeutic agent is substantially released from the vivo polymerizedbiocompatible retinal patch within 180 days. In certain embodiments, thetherapeutic agent is substantially released from the vivo polymerizedbiocompatible retinal patch within 24 hours. In some embodiments, therelease of the therapeutic agent is determined by the composition of thein vivo polymerized biocompatible retinal patch. In certain embodiments,the therapeutic agent is released while the in vivo polymerizedbiocompatible retinal patch degrades. In some embodiments, the releaseof the therapeutic agent is essentially inhibited until a time that thein vivo polymerized biocompatible retinal patch starts to degrade. Incertain embodiments, at least a portion of the therapeutic agent isreleased before the time that the in vivo polymerized biocompatibleretinal patch starts to degrade. In some embodiments, the time the invivo polymerized biocompatible retinal patch starts to degrade is longerthe higher a degree of cross-linking of the in vivo polymerizedbiocompatible retinal patch. In certain embodiments, the time the invivo polymerized biocompatible retinal patch starts to degrade isshorter the higher a concentration of ester groups in the first orsecond compound.

Also provided herein is a method of treating retinal detachment, aretinal tear, or a retinal hole, comprising delivering an in vivogelling ophthalmic pre-formulation to a site of a retinal tear in ahuman eye, the in vivo gelling ophthalmic pre-formulation comprising:(a) at least one first compound comprising more than one nucleophilicgroup; (b) at least one second compound comprising more than oneelectrophilic group; (c) an aqueous buffer in the pH range of about 6.0to about 8.5; and (d) a viscosity enhancer; wherein the in vivo gellingophthalmic formulation at least in part polymerizes and/or gels at thesite of the retinal tear in the human eye to form a biocompatibleretinal patch. In some embodiments, the biocompatible retinal patch atleast partially adheres to the site of the retinal tear. In certainembodiments, the biocompatible retinal patch closes the site of aretinal tear.

In some embodiments of the method, the in vivo gelling ophthalmicpre-formulation further comprises a therapeutic agent. In certainembodiments, the viscosity enhancer is selected fromhydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose,polyvinyl alcohol, or polyvinylpyrrolidone.

In certain embodiments of the method, the nucleophilic group is a thiolor amino group. In some embodiments, the first compound is a glycol,trimethylolpropane, pentaerythritol, hexaglycerol, or tripentaerythritolderivative. In certain embodiments, the first compound further comprisesone or more polyethylene glycol sections. In some embodiments, the firstcompound is a pentaerythritol or hexaglycerol derivative. In certainembodiments, the first compound is selected from the group consisting ofethoxylated pentaerythritol ethylamine ether, ethoxylatedpentaerythritol propylamine ether, ethoxylated pentaerythritol aminoacetate, ethoxylated hexaglycerol ethylamine ether, ethoxylatedhexaglycerol propylamine ether, and ethoxylated hexaglycerol aminoacetate. In some embodiments, the first compound is selected from thegroup consisting of trimethylolpropane trimercaptoacetate,trimethylolpropane tri-3-mercaptopropionate, pentaerythritoltetramercaptoacetate, pentaerythritol tetra-3-mercaptopropionate,ethoxylated trimethylolpropane trimercaptoacetate, ethoxylatedtrimethylolpropane tri-3-mercaptopropionate, ethoxylated pentaerythritoltetramercaptoacetate, and ethoxylated trimethylolpropanetri-3-mercaptopropionate. In certain embodiments, the molecular weightof the first compound is between about 100 and 100000. In someembodiments, the first compound is water soluble.

In some embodiments of the method, the electrophilic group is anepoxide, N-succinimidyl succinate, N-succinimidyl glutarate,N-succinimidyl succinamide or N-succinimidyl glutaramide. In certainembodiments, the second compound is a trimethylolpropane, glycerol,diglycerol, pentaerythritol, sorbitol, hexaglycerol, tripentaerythritol,or polyglycerol derivative. In some embodiments, the second compoundfurther comprises one or more polyethylene glycol sections. In certainembodiments, the second compound is a trimethylolpropane,pentaerythritol, or hexaglycerol derivative. In some embodiments, thesecond compound is selected from the group consisting of ethoxylatedpentaerythritol succinimidyl succinate, ethoxylated pentaerythritolsuccinimidyl glutarate, ethoxylated pentaerythritol succinimidylglutaramide, ethoxylated hexaglycerol succinimidyl succinate,ethoxylated hexaglycerol succinimidyl glutarate, and ethoxylatedhexaglycerol succinimidyl glutaramide. In certain embodiment, the secondcompound is selected from the group consisting of sorbitol polyglycidylether, polyglycerol polyglycidyl ether, diglycerol polyglycidyl ether,glycerol polyglycidyl ether, and trimethylolpropane polyglycidyl ether.In some embodiments, the molecular weight of the second compound isbetween about 10000 and 100000. In certain embodiments, the secondcompound is water soluble.

In certain embodiments of the method, the gelling time of thebiocompatible retinal patch is controlled by the pH of the aqueousbuffer, the type of the buffer, the concentration of the buffer, theconcentration of the first compound and/or the second compound in thebuffer, or the nature of the electrophilic groups. In some embodiments,the gelling time is between about 20 seconds and 10 minutes. In certainembodiments, the pH of the aqueous buffer is from about 8. In someembodiments, the in vivo gelling ophthalmic pre-formulation gels at apredetermined time to form the biocompatible retinal patch.

In some embodiments of the method, the biocompatible retinal patch is abioabsorbable polymer. In certain embodiments of the method, thebiocompatible retinal patch is bioabsorbed within about 1 to 70 days. Insome embodiments of the method, the biocompatible retinal patch issubstantially non-bioabsorbable.

In certain embodiments of the method, the therapeutic agent is releasedfrom the biocompatible retinal patch through diffusion, osmosis,degradation of the biocompatible retinal patch, or any combinationthereof. In some embodiments, the therapeutic agent is initiallyreleased from the biocompatible retinal patch through diffusion andlater released through degradation of the biocompatible retinal patch.In certain embodiments, the therapeutic agent is substantially releasedfrom the biocompatible retinal patch within 180 days. In someembodiments, the therapeutic agent is substantially released from thebiocompatible retinal patch within 14 days. In certain embodiments, thetherapeutic agent is substantially released from the biocompatibleretinal patch within 24 hours. In some embodiments, the therapeuticagent is substantially released from the biocompatible retinal patchwithin one hour. In certain embodiments, the first compound and thesecond compound do not react with the therapeutic agent during formationof the biocompatible retinal patch. In some embodiments, thebiocompatible retinal patch interacts with the therapeutic agent, andmore than 10% of the therapeutic agent is released through degradationof the biocompatible retinal patch. In certain embodiments, more than30% of the therapeutic agent is released through degradation of thebiocompatible retinal patch. In some embodiments, the biocompatibleretinal patch interacts with the therapeutic agent by forming covalentbonds between the biocompatible retinal patch and the therapeutic agent.In certain embodiments, the biocompatible retinal patch interacts withthe therapeutic agent by forming a non-covalent bond between thebiocompatible retinal patch and the therapeutic agent. In someembodiments, the therapeutic agent is released while the biocompatibleretinal patch degrades. In certain embodiments, the release of thetherapeutic agent is essentially inhibited until a time that thebiocompatible retinal patch starts to degrade. In some embodiments, thetime the biocompatible retinal patch starts to degrade is longer thehigher a degree of cross-linking of the biocompatible retinal patch. Incertain embodiments, the time the biocompatible retinal patch starts todegrade is shorter the higher a concentration of ester groups in thefirst or second compound.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 shows the effect of pH on monomer addition to the 0.10 Mphosphate reaction buffer for two formulations: 1) 8ARM-20k-NH2 &4ARM-20k-SGA at 4.8% solution with 0.3% HPMC; 2)8ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA at 4.8% solution with0.3% HPMC.

FIG. 2 shows the effect of reaction buffer pH on gel times for8ARM-20k-NH2 & 8ARM-15k-SG polymer formulation at 5% solution using 0.05M phosphate buffer.

FIG. 3 shows the effect of reaction buffer phosphate concentration ongel times for 8ARM-20k-NH2 & 8ARM-15k-SG polymer formulation at 5%solution using phosphate buffer at pH 7.4.

FIG. 4 shows the effect of polymer concentration on gel times using a0.05 M phosphate buffer at pH 7.4.

FIG. 5 shows the effect of temperature on gel times for twoformulations: 1) 8ARM-20k-NH2 & 4ARM-20k-SGA at 4.8% solution with 0.3%HPMC; 2) 8ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA at 4.8%solution with 0.3% HPMC.

FIG. 6 shows the viscosity of the polymer solution up to the gel pointas a function of the % of time to the gel point.

FIG. 7 shows the bulk monomer stability over approximately 56 weeks forthe formulation involving 8ARM-20k-NH2, 4ARM-20k-AA, and 8ARM-15k-SG atan amine to ester molar ratio of 1 to 1. The % solution of polymer was5%±0.5%. Different reaction buffers were used over time, but thecomposition was typically 0.058 M phosphate at a pH of 7.4±0.1.

FIG. 8 shows the effect of addition of degradable acetate amine8ARM-20k-AA or 4ARM-20k-AA on degradation times. Degradations occurredin phosphate buffered saline (PBS) at 37° C.

FIG. 9 shows the effect of degradation buffer pH on degradation times.Degradations occurred at 37° C.

FIGS. 10A, 10B, 10C, and 10D show several photos depicting the8ARM-20k-NH2 & 4ARM-20k-SGA at 4.8% solution with 0.3% HPMC formulationas an example of a smooth degradation process. The polymer was createdin a cylindrical shape with red food dye for visualization purposes. Theinitial state of the polymer in degradation buffer is shown (FIG. 10A).After several days, the polymer swelled, but retained its shape (FIG.10B). As the degradation point is approached, the polymer became softand lost its shape (FIG. 10C). Finally, the polymer degraded into thesolution (FIG. 10D).

FIGS. 11A and 11B show photos depicting two different formulations asexamples of fragmenting degradation processes. The polymer was createdin a cylindrical shape with red food dye for visualization purposes. The8ARM-20k-AA/8ARM-20k-NH2 (60/40) & 4ARM-20k-SGA at 4.8% solution with0.3% HPMC is shown near the degradation point (FIG. 11A). The4ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA at 4.8% solution with0.3% HPMC is shown near the degradation point (FIG. 11B).

FIGS. 12A, 12B, 12C, 12D, and 12E show the effect of polymer solution ongel time (FIG. 12A), firmness (FIG. 12B), tack (FIG. 12C), elasticmodulus (FIG. 12D), and swelling (FIG. 12E) for the formulation:8ARM-20k-AA/8ARM-20k-NH2 (75/25) & 4ARM-20k-SGA with 0.3% HPMC. Theerror bars represent the standard deviations of 3 samples.

FIG. 13 shows the effect of hypromellose (HPMC) addition at 0, 0.3 and1.0% to the polymer formulations on swelling.

FIG. 14 shows the specific gravity of polymer solutions in relation topure water.

FIG. 15 shows the effect of barium sulfate (BaSO₄) on the solutionviscosity for concentrations of 0.0, 1.0, 2.0, 5.0 and 10.0% (w/v).

FIG. 16 shows a sample plot generated by the Texture Analyzer Exponentsoftware running the firmness test. The peak force was recorded as thepolymer firmness, which represents the point where the targetpenetration depth of 4 mm has been reached by the probe.

FIG. 17 shows a sample plot generated by the Texture Analyzer Exponentsoftware running the elastic modulus test under compression. The moduluswas calculated from the initial slope of the curve up to 10% of themaximum compression stress.

FIG. 18 shows an exemplary plot generated by the Texture AnalyzerExponent software running the adhesion test. A contact force of 100.0 gwas applied for 10 seconds. The tack was measured as the peak forceafter lifting the probe from the sample. The adhesion energy or the workof adhesion was calculated as the area under the curve representing thetack force (points 1 to 2). The stringiness was defined as the distancetraveled by the probe while influencing the tack force (points 1 and 2).

FIGS. 19A and 19B show the effect of hypromellose (HPMC) addition at 0,0.3 and 1.0% to the polymer formulations on firmness (FIG. 19A). Effectof degradable acetate amine 8ARM-20k-AA addition at 0, 70 and 100% tothe polymer formulations on firmness (FIG. 19B).

FIGS. 20A and 20B show the effect of hypromellose (HPMC) addition at 0,0.3 and 1.0% to the polymer formulations on the elastic modulus (FIG.20A) and shows the effect Effect of degradable acetate amine 8ARM-20k-AAaddition at 0, 70 and 100% to the polymer formulations on the elasticmodulus (FIG. 20B).

FIGS. 21A, 21B, 21C, and 21D show a comparison of the firmness (FIG.21A), tack (FIG. 21B), adhesion energy (FIG. 21C) and stringiness (FIG.21D) of the general polymer formulation: 8ARM-20k-AA/8ARM-20k-NH2 (x/y)& 4ARM-20k-SGA at 4.8% solution with 0.3% HPMC. The measured values fora Post-It™ note are included as a reference.

FIG. 22 shows the firmness vs. degradation time plotted as percentagesfor the polymer formulation: 8ARM-20k-AA/8ARM-20k-NH2 (70/30) &4ARM-20k-SGA at 4.8% solution with 0.3% HPMC. The error bars representthe standard deviations of 3 samples. The degradation time for thepolymer was 18 days.

FIGS. 23A and 23B show the general assembly of directly connectedsyringes for use in a kit.

FIG. 24 shows the general design of a mixing assembly (red food dye wasadded for clarity purposes).

DETAILED DESCRIPTION OF THE INVENTION

The current method for treating retinal detachment involves surgeryfollowed by suturing or laser treatment followed by filling the eyeinterior with an inert gas or silicon oil. The post operation results inthe intraocular pressure inside and there is no vision for 2-4 weeks. Iffilled with the inert gas, the travel is also not possible during thisperiod due to the possible change in the atmospheric pressure at the newlocation. However, if the cavity is filled with silicone oil, thenanother surgery is required to remove the silicon oil after the healingprocess is complete. In addition, the current retinal patches aredifficult to make stay in place while the current commercially availableadhesives do not bond well to the retina and are difficult to deliver tothe site.

Therefore, herein is provided a family of in vivo gelling biocompatibleophthalmic pre-formulations that can be injected at or near the affectedarea using a narrow bore needles and that form biocompatible retinalpatches. Once the target location has been identified inside the eye, anexact volume of the reacting mixture is injected inside. Once on site,the liquid wets the surface of the retina and fills the hole. After apreset time, the liquid turns into a solid and bonds the two layerstogether and attaches the retina and also fills the hole forming aretinal patch. The viscosity of the formulations is controlled such thatthe liquid remains localized at the target site. In some embodiments,the in vivo gelling biocompatible ophthalmic pre-formulation alsoadheres to the site in the eye. In certain embodiment, the viscosity andstickiness of the in vivo gelling ophthalmic pre-formulation is suitablefor easy delivery to the site through a narrow bore needle, while at thesame time staying in place at the site of the retinal tear and adheringto the tissue surrounding the retinal tear. In some embodiments, the invivo gelling ophthalmic pre-formulations comprise viscosity enhancers toensure that the pre-formulation remains at and/or adheres to the targetsite in the eye during the gelling process. The physicochemicalproperties of the biopolymer are matched with the surrounding tissuesand the retina, so that there is little change in pressure from thenatural atmosphere.

In further embodiments, an in vivo gelling ophthalmic pre-formulation toform a biocompatible retinal patch enables the administration ofmedication directly to the vitreous. The polymer starts out as a liquidpre-formulation and is delivered, together with one or more optionaltherapeutic agents, to the site of a disease using minimally invasivetechniques. Once in the eye, the liquid pre-formulation polymerizes intoa solid hydrogel that in some instances adheres to the tissue and keepsthe polymer/drug combination at the site of the disease. In someinstances, polymerization and degradation times are controlled byvarying the composition of the monomers and buffers allowing for theappropriate application and placement of the hydrogel polymer. In someembodiments, the drug is released in a precise and consistent manner. Incertain instances, the biocompatible hydrogel polymer is bioabsorbedover a defined period of time. In some embodiments, the biocompatiblehydrogel polymer provides the sustained release of a therapeutic agentat a target site. In certain embodiments, the sustained and controlledrelease reduces the systemic exposure to the therapeutic agent. Thecontrolled gelling and biodegradation allows the use of thebiocompatible hydrogel polymer to deliver one or more therapeutic agentsdirectly to the tissue affected by a disease, thereby minimizingsystemic exposure to the therapeutic agent.

In some instances, the therapeutic agent is released from thebiocompatible hydrogel polymer over an extended period of time. Incertain instances, delivery of the therapeutic agent in a biocompatiblehydrogel polymer provides a depot of the therapeutic agent (e.g., underthe skin), wherein the depot releases the therapeutic agent over anextended period of time (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6days, 7 days, 10, days, 14 days, 3 week, 4 week). In some instances, thebiocompatible hydrogel polymer releases the therapeutic agent after adelay as a delayed burst.

In some embodiments, an in vivo gelling ophthalmic pre-formulationcompletely replaces the vitreous humour of an eye. In certainembodiments, the in vivo gelling ophthalmic pre-formulation partiallyreplaces the vitreous humour of an eye.

The in vivo gelling ophthalmic pre-formulations are useful to treatretinal detachment and other conditions of the eye. Delivering an invivo gelling ophthalmic pre-formulation to the site of a retinal tear inan eye, the pre-formulation at least in part polymerizes and/or gels atthe site of the retinal tear in the eye and forms a retinal patch. Theretinal patch at least partially adheres to the retinal tear and closesthe hole treating the retinal detachment and allowing it to heal.

Furthermore, the in vivo gelling ophthalmic pre-formulations are usefulto deliver therapeutic agents to the inside of the eye to treatconditions of the eye, for example age-related macular degeneration,diabetic retinopathy, cataract, intra-ocular pressure (glaucoma), orproliferative vitreoretinopathy.

Exemplary Hydrogel Components

Provided herein are in vivo gelling ophthalmic pre-formulations,comprising at least one first compound comprising more than onenucleophilic group, at least one second compound comprising more thanone electrophilic group, an aqueous buffer in the pH range of about 5.0to about 9.5, and optionally one or more therapeutic agents. In certainembodiments, the in vivo gelling ophthalmic pre-formulation forms abiocompatible hydrogel polymer at a target site in a human body bymixing the at least one first compound, the at least one secondcompound, and the optional therapeutic agent in the aqueous buffer anddelivering the mixture to the target site such that the biocompatiblehydrogel polymer at least in part polymerizes and/or gels at the targetsite. In some embodiments, the biocompatible hydrogel polymer is formedfollowing mixing the first compound and the second compound in theaqueous buffer; and wherein the biocompatible hydrogel polymer gels at atarget site. In certain embodiments, mixing the first compound, thesecond compound, and the optional therapeutic agent in the aqueousbuffer and delivering the mixture to a target site in the human bodygenerates the in vivo gelling ophthalmic pre-formulation such that thein vivo gelling ophthalmic pre-formulation at least in part polymerizesand/or gels at the target site to form a biocompatible hydrogel polymer.

In some embodiments, the first or second compound comprising more thanone nucleophilic or electrophilic group are polyol derivatives. Incertain embodiments, the first or second compound is a dendritic polyolderivative. In some embodiments, the first or second compound is aglycol, trimethylolpropane, glycerol, diglycerol, pentaerythritiol,sorbitol, hexaglycerol, tripentaerythritol, or polyglycerol derivative.In certain embodiments, the first or second compound is a glycol,trimethylolpropane, pentaerythritol, hexaglycerol, or tripentaerythritolderivative. In some embodiments, the first or second compound is atrimethylolpropane, glycerol, diglycerol, pentaerythritiol, sorbitol,hexaglycerol, tripentaerythritol, or polyglycerol derivative. In someembodiments, the first or second compound is a pentaerythritol,di-pentaerythritol, or tripentaerythritol derivative. In certainembodiments, the first or second compound is a hexaglycerol(2-ethyl-2-(hydroxymethyl)-1,3-propanediol, trimethylolpropane)derivative. In some embodiments, the first or second compound is asorbitol derivative. In certain embodiments, the first or secondcompound is a glycol, propyleneglycol, glycerin, diglycerin, orpolyglycerin derivative.

In some embodiments, the first and/or second compound further comprisespolyethylene glycol (PEG) chains comprising one to 200 ethylene glycolsubunits. In certain embodiments, the first and/or second compoundfurther comprises polypropylene glycol (PPG) chains comprising one to200 propylene glycol subunits. The PEG or PPG chains extending from thepolyols are the “arms” linking the polyol core to the nucleophilic orelectrophilic groups.

Exemplary Nucleophilic Monomers

The in vivo gelling ophthalmic pre-formulation comprises at least onefirst compound comprising more than one nucleophilic group. In someembodiments, the nucleophilic group is a hydroxyl, thiol, or aminogroup. In preferred embodiments, the nucleophilic group is a thiol oramino group.

In certain embodiments, the nucleophilic group is connected to thepolyol derivative through a suitable linker. Suitable linkers include,but are not limited to, esters (e.g., acetates) or ethers. In someinstances, monomers comprising ester linkers are more susceptible tobiodegradation. Examples of linkers comprising a nucleophilic groupinclude, but are not limited to, mercaptoacetate, aminoacetate (glycin)and other amino acid esters (e.g., alanine, β-alanine, lysine,ornithine), 3-mercaptopropionate, ethylamine ether, or propylamineether. In some embodiments, the polyol core derivative is bound to apolyethylene glycol or polypropylene glycol subunit, which is connectedto the linker comprising the nucleophilic group. The molecular weight ofthe first compound (the nucleophilic monomer) is about 100 to 100000. Incertain embodiments, the molecular weight of a first compound (anucleophilic monomer) is about 100, about 500, about 1000, about 2000,about 3000, about 4000, about 5000, about 6000, about 7000, about 8000,about 9000, about 10000, about 12000, about 15000, about 20000, about25000, about 30000, about 35000, about 40000, about 50000, about 60000,about 70000, about 80000, about 90000, or about 100000. In certainembodiments, the molecular weight of a second compound is about 500 to40000. In some embodiments, the molecular weight of a first compound isabout 500 to 2000. In certain embodiments, the molecular weight of afirst compound is about 15000 to about 40000. In some embodiments, thefirst compound is water soluble.

Examples of the construction of monomers comprising more than onenucleophilic group are shown below with a trimethylolpropane orpentaerythritol core polyol. The compounds shown have thiol or amineelectrophilic groups that are connected to variable lengths PEG subunitthrough acetate, propionate or ethyl ether linkers (e.g., structuresbelow of ETTMP (A; n=1), 4ARM-PEG-NH2 (B; n=1), and 4ARM-PEG-AA (C;n=1)). Monomers using other polyol cores are constructed in a similarway.

Suitable first compounds comprising a nucleophilic group (used in theamine-ester chemistry) include, but are not limited to, pentaerythritolpolyethylene glycol amine (4ARM-PEG-NH2) (molecular weight selected fromabout 5000 to about 40000, e.g., 5000, 10000, or 20000), pentaerythritolpolyethylene glycol amino acetate (4ARM-PEG-AA) (molecular weightselected from about 5000 to about 40000, e.g., 5000, 10000, or 20000),hexaglycerin polyethylene glycol amine (8ARM-PEG-NH2) (molecular weightselected from about 5000 to about 40000, e.g., 10000, 20000, or 40000),or tripentaerythritol glycol amine (8ARM(TP)-PEG-NH2) (molecular weightselected from about 5000 to about 40000, e.g., 10000, 20000, or 40000).Within this class of compounds, 4(or 8)ARM-PEG-AA comprises ester (oracetate) groups while the 4(or 8)ARM-PEG-NH2 monomers do not compriseester (or acetate) groups.

Other suitable first compounds comprising a nucleophilic group (used inthe thiol-ester chemistry) include, but not limited to, glycoldimercaptoacetate (THIOCURE® GDMA), trimethylolpropanetrimercaptoacetate (THIOCURE® TMPMA), pentaerythritoltetramercaptoacetate (THIOCURE® PETMA), glycol di-3-mercaptopropionate(THIOCURE® GDMP), trimethylolpropane tri-3-mercaptopropionate (THIOCURE®TMPMP), pentaerythritol tetra-3-mercaptopropionate (THIOCURE® PETMP),polyol-3-mercaptopropionates, polyester-3-mercaptopropionates,propyleneglycol 3-mercaptopropionate (THIOCURE® PPGMP 800),propyleneglycol 3-mercaptopropionate (THIOCURE® PPGMP 2200), ethoxylatedtrimethylolpropane tri-3-mercaptopropionate (THIOCURE® ETTMP-700), andethoxylated trimethylolpropane tri-3-mercaptopropionate (THIOCURE®ETTMP-1300).

Exemplary Electrophilic Monomers

The in vivo gelling ophthalmic pre-formulation comprises at least onefirst compound comprising more than one electrophilic group. In someembodiments, the electrophilic group is an epoxide, maleimide,succinimidyl, or an alpha-beta unsaturated ester. In preferredembodiments, the electrophilic group is an epoxide or succinimidyl.

In certain embodiments, the electrophilic group is connected to thepolyol derivative through a suitable linker. Suitable linkers include,but are not limited to, esters, amides, or ethers. In some instances,monomers comprising ester linkers are more susceptible tobiodegradation. Examples of linkers comprising an electrophilic groupinclude, but are not limited to, succinimidyl succinate, succinimidylglutarate, succinimidyl succinamide, succinimidyl glutaramide, orglycidyl ether. In some embodiments, the polyol core derivative is boundto a polyethylene glycol or polypropylene glycol subunit, which isconnected to the linker comprising the electrophilic group. Themolecular weight of the second compound (the electophilic monomer) isabout 100 to 100000. In certain embodiments, the molecular weight of asecond compound (an electophilic monomer) is about 100, about 500, about1000, about 2000, about 3000, about 4000, about 5000, about 6000, about7000, about 8000, about 9000, about 10000, about 12000, about 15000,about 20000, about 25000, about 30000, about 35000, about 40000, about50000, about 60000, about 70000, about 80000, about 90000, or about100000. In certain embodiments, the molecular weight of a secondcompound is about 500 to 40000. In some embodiments, the molecularweight of a second compound is about 500 to 2000. In certainembodiments, the molecular weight of a second compound is about 15000 toabout 40000. In some embodiments, the second compound is water soluble.

Examples of the construction of monomers comprising more than oneelectrophilic group are shown below with a pentaerythritol core polyol.The compounds shown have a succinimidyl electrophilic group, a glutarateor glutaramide linker, and a variable lengths PEG subunit (e.g.,structures below of 4ARM-PEG-SG (D; n=3) and 4ARM-PEG-SGA (E; n=3)).Monomers using other polyol cores or different linkers (e.g., succinate(SS) or succinamide (SSA) are constructed in a similar way.

Suitable second compounds comprising an electrophilic group include, butare not limited to, pentaerythritol polyethylene glycol maleimide(4ARM-PEG-MAL) (molecular weight selected from about 5000 to about40000, e.g., 10000 or 20000), pentaerythritol polyethylene glycolsuccinimidyl succinate (4ARM-PEG-SS) (molecular weight selected fromabout 5000 to about 40000, e.g., 10000 or 20000), pentaerythritolpolyethylene glycol succinimidyl glutarate (4ARM-PEG-SG) (molecularweight selected from about 5000 to about 40000, e.g., 10000 or 20000),pentaerythritol polyethylene glycol succinimidyl glutaramide(4ARM-PEG-SGA) (molecular weight selected from about 5000 to about40000, e.g., 10000 or 20000), hexaglycerin polyethylene glycolsuccinimidyl succinate (8ARM-PEG-SS) (molecular weight selected fromabout 5000 to about 40000, e.g., 10000 or 20000), hexaglycerinpolyethylene glycol succinimidyl glutarate (8ARM-PEG-SG) (molecularweight selected from about 5000 to about 40000, e.g., 10000, 15000,20000, or 40000), hexaglycerin polyethylene glycol succinimidylglutaramide (8ARM-PEG-SGA) (molecular weight selected from about 5000 toabout 40000, e.g., 10000, 15000, 20000, or 40000), tripentaerythritolpolyethylene glycol succinimidyl succinate (8ARM(TP)-PEG-SS) (molecularweight selected from about 5000 to about 40000, e.g., 10000 or 20000),tripentaerythritol polyethylene glycol succinimidyl glutarate(8ARM(TP)-PEG-SG) (molecular weight selected from about 5000 to about40000, e.g., 10000, 15000, 20000, or 40000), or tripentaerythritolpolyethylene glycol succinimidyl glutaramide (8ARM(TP)-PEG-SGA)(molecular weight selected from about 5000 to about 40000, e.g., 10000,15000, 20000, or 40000). The 4(or 8)ARM-PEG-SG monomers comprise estergroups, while the 4(or 8)ARM-PEG-SGA monomers do not comprise estergroups.

Other suitable second compounds comprising an electrophilic group aresorbitol polyglycidyl ethers, including, but not limited to, sorbitolpolyglycidyl ether (DENACOL® EX-611), sorbitol polyglycidyl ether(DENACOL® EX-612), sorbitol polyglycidyl ether (DENACOL® EX-614),sorbitol polyglycidyl ether (DENACOL® EX-614 B), polyglycerolpolyglycidyl ether (DENACOL® EX-512), polyglycerol polyglycidyl ether(DENACOL® EX-521), diglycerol polyglycidyl ether (DENACOL® EX-421),glycerol polyglycidyl ether (DENACOL® EX-313), glycerol polyglycidylether (DENACOL® EX-313), trimethylolpropane polyglycidyl ether (DENACOL®EX-321), sorbitol polyglycidyl ether (DENACOL® EJ-190).

Viscosity Enhancer

The in vivo gelling ophthalmic pre-formulation, the biocompatibleretinal patch, and the in vivo polymerized biocompatible retinal patchcomprise a viscosity enhancer. In some instances, the viscosity enhancerincreases the viscosity of the pre-formulation, preventing thepre-formulation from spreading and allowing to pre-formulation to stayat the target site. Viscosity enhancers include, but are not limited to,acacia, agar, alginic acid, bentonite, carbomers, carboxymethylcellulosecalcium, carboxymethylcellulose sodium, carrageenan, ceratonia,cetostearyl alcohol, chitosan, colloidal silicon dioxide,cyclomethicone, ethylcellulose, gelatin, glycerin, glyceryl behenate,guar gum, hectorite, hydrogenated vegetable oil type I, hydroxyethylcellulose, hydroxyethylmethyl cellulose, hydroxypropyl cellulose,hydroxypropyl starch, hypromellose, magnesium aluminum silicate,maltodextrin, methylcellulose, polydextrose, polyethylene glycol,poly(methylvinyl ether/maleic anhydride), polyvinyl acetate phthalate,polyvinyl alcohol, potassium chloride, povidone, propylene glycolalginate, saponite, sodium alginate, sodium chloride, stearyl alcohol,sucrose, sulfobutylether (3-cyclodextrin, tragacanth, xanthan gum, andderivatives and mixtures thereof. In certain embodiments, the viscosityenhancer is selected from hydroxyethylcellulose,hydroxypropylmethylcellulose, methylcellulose, polyvinyl alcohol, orpolyvinylpyrrolidone. In some embodiments, the viscosity enhancer ismethylcellulose or hydroxypropylmethylcellulose. In preferredembodiments, the viscosity enhancer is hydroxypropylmethylcellulose.

In some embodiments, the viscosity enhancer is a bioadhesive orcomprises a bioadhesive polymer. In certain instances, a bioadhesive isany adhesive that interfaces with living tissue and/or biological fluid.Bioadhesive polymers include, but are not limited to,hydroxypropylmethylcellulose, hydroxyethyl cellulose, hydroxypropylcellulose, carboxymethyl cellulose, polyvinyl alcohol, sodiumhyaluronate, chitosan, alginate, xanthum gum, acrylic polymers (e.g.,carbomer, polycarbophil), and derivatives and mixtures thereof.

In some embodiments, the concentration of the viscosity enhancer in thebuffer ranges from 0.1 to 20%. In certain embodiments, the concentrationof the viscosity enhancer in the buffer ranges from 5 to 20%. In otherembodiments, the concentration of the viscosity enhancer in the bufferranges from 0.1 to 2%. In specific embodiments, the concentration of theviscosity enhancer in the buffer rangers from 0.1 to 0.5%. In someembodiments, the concentration of the viscosity enhancer is less than20%, less than 15%, less than 10%, less than 9%, less than 8%, less than7%, less than 6%, less than 5%, less than 4%, less than 3%, less than2%, less than 1.8%, less than 1.6%, less than 1.5%, less than 1.4%, lessthan 1.2%, less than 1%, less than 0.9%, less than 0.8%, less than 0.7%,less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, lessthan 0.2%, or less than 0.1%. In some embodiments, the concentration ofthe viscosity enhancer is at least 10%, at least 8%, at least 6%, atleast 5%, at least 4%, at least 2%, at least 1.8%, at least 1.6%, atleast 1.5%, at least 1.4%, at least 1.2%, at least 1%, at least 0.9%, atleast 0.8%, at least 0.7%, at least 0.6%, at least 0.5%, at least 0.4%,at least 0.3%, at least 0.2%, or at least 0.1%. In some embodiments, theconcentration of the viscosity enhancer is about 20%, about 15%, about10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about3%, about 2%, about 1.8%, about 1.6%, about 1.5%, about 1.4%, about1.2%, about 1%, about 0.9%, about 0.8%, about 0.7%, about 0.6%, about0.5%, about 0.4%, about 0.3%, about 0.2%, or about 0.1%.

In certain embodiments, the viscosity of the in vivo gelling ophthalmicpre-formulation is less than 4000 cP, less than 2000 cP, less than 1000cP, less than 800 cP, less than 600 cP, less than 500 cP, less than 400cP, less than 200 cP, less than 100 cP, less than 80 cP, less than 60cP, less than 50 cP, less than 40 cP, less than 20 cP, less than 10 cP,less than 8 cP, less than 6 cP, less than 5 cP, less than 4 cP, lessthan 3 cP, less than 2 cP, less than 1 cP. In some embodiments, theviscosity of the in vivo gelling ophthalmic pre-formulation is at least4000 cP, at least 2000 cP, at least 1000 cP, at least 800 cP, at least600 cP, at least 500 cP, at least 400 cP, at least 200 cP, at least 100cP, at least 80 cP, at least 60 cP, at least 50 cP, at least 40 cP, atleast 20 cP, at least 10 cP, at least 8 cP, at least 6 cP, at least 5cP, at least 4 cP, at least 3 cP, at least 2 cP, at least 1 cP. Incertain embodiments, the viscosity of the in vivo gelling ophthalmicpre-formulation is about 4000 cP, about 2000 cP, about 1000 cP, about800 cP, about 600 cP, about 500 cP, about 400 cP, about 200 cP, about100 cP, about 80 cP, about 60 cP, about 50 cP, about 40 cP, about 20 cP,about 10 cP, about 8 cP, about 6 cP, about 5 cP, about 4 cP, about 3 cP,about 2 cP, about 1 cP. In some embodiments, the viscosity of the invivo gelling ophthalmic pre-formulation is between about 5 cP and 50 cP.In certain embodiments, the viscosity of the in vivo gelling ophthalmicpre-formulation is between about 5 cP and 500 cP.

Formation of Hydrogels

In certain embodiments, the first and second compounds comprising morethan one nucleophilic or more than one electrophilic group safelyundergo polymerization at a target site inside a mammalian body, forinstance in the eye, for example at the site of a retinal tear. Incertain embodiments, the in vivo gelling ophthalmic pre-formulationreplaces, partially or complete, the vitreous in the eye. In someembodiments, the first compound and the second compound are monomersforming a polymer through the reaction of a nucleophilic group in thefirst compound with the electrophilic group in the second compound. Incertain embodiments, the monomers are polymerized at a predeterminedtime. In some embodiments, the monomers are polymerized under mild andnearly neutral pH conditions. In certain embodiments, the hydrogelpolymer does not change volume after curing.

In some embodiments, the first and second compound react to form amide,thioester, or thioether bonds. When a thiol nucleophile reacts with asuccinimidyl electrophile, a thioester is formed. When an aminonucleophile reacts with a succinimidyl electrophile, an amide is formed.

In some embodiments, one or more first compounds comprising an aminogroup react with one or more second compounds comprising a succinimidylester group to form amide linked first and second monomer units. Incertain embodiments, one or more first compounds comprising a thiolgroup react with one or more second compounds comprising a succinimidylester group to form thioester linked first and second monomer units. Insome embodiments, one or more first compounds comprising an amino groupreact with one or more second compounds comprising an epoxide group tofrom amine linked first and second monomer units. In certainembodiments, one or more first compounds comprising a thiol group reactwith one or more second compounds comprising an epoxide group to formthioether linked first and second monomer units.

In some embodiments, a first compound is mixed with a different firstcompound before addition to one or more second compounds. In otherembodiments, a second compound is mixed with a different second compoundbefore addition to one or more first compounds. In certain embodiments,the properties of the in vivo gelling ophthalmic pre-formulation and thebiocompatible hydrogel polymer are controlled by the properties of theat least one first and at least one second monomer mixture.

In some embodiments, one first compound is used in the biocompatiblehydrogel polymer. In certain embodiments, two different first compoundsare mixed and used in the biocompatible hydrogel polymer. In someembodiments, three different first compounds are mixed and used in thebiocompatible hydrogel polymer. In certain embodiments, four or moredifferent first compounds are mixed and used in the biocompatiblehydrogel polymer.

In some embodiments, one second compound is used in the biocompatiblehydrogel polymer. In certain embodiments, two different second compoundsare mixed and used in the biocompatible hydrogel polymer. In someembodiments, three different second compounds are mixed and used in thebiocompatible hydrogel polymer. In certain embodiments, four or moredifferent second compounds are mixed and used in the biocompatiblehydrogel polymer.

In some embodiments, a first compound comprising ether linkages to thenucleophilic group are mixed with a different first compound comprisingester linkages to the nucleophilic group. This allows the control of theconcentration of ester groups in the resulting biocompatible hydrogelpolymer. In certain embodiments, a second compound comprising esterlinkages to the electrophilic group are mixed with a different secondcompound comprising ether linkages to the electrophilic group. In someembodiments, a second compound comprising ester linkages to theelectrophilic group are mixed with a different second compoundcomprising amide linkages to the electrophilic group. In certainembodiments, a second compound comprising amide linkages to theelectrophilic group are mixed with a different second compoundcomprising ether linkages to the electrophilic group.

In some embodiments, a first compound comprising an aminoacetatenucleophile is mixed with a different first compound comprising anethylamine ether nucleophile at a specified molar ratio (x/y). Incertain embodiments, the molar ratio (x/y) is 5/95, 10/90, 15/85, 20/80,25/75, 30/70, 35/65, 40/60, 45/55, 50/50, 55/45, 60/40, 65/35, 70/30,75/25, 80/20, 85/15, 90/10, or 95/5. In certain embodiments, the mixtureof two first compounds is mixed with one or more second compounds at amolar amount equivalent to the sum of x and y. In some embodiments, theratio of the combined molar amount of the two first compounds to themolar amount of the second compound is not equivalent.

In some embodiments, the molar ratio of the combined molar amount of thefirst compounds to the combined molar amount of the second compounds isgreater than 1. In certain embodiments, the molar ratio of the combinedmolar amount of the first compounds to the combined molar amount of thesecond compounds is less than 1. In some embodiments, the molar ratio ofthe combined molar amount of the first compounds to the combined molaramount of the second compounds is about 1. In certain embodiments, themolar ratio of the combined molar amount of the first compounds to thecombined molar amount of the second compounds is about 10:1, about 9:1,about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about2:1, or about 1:1. In some embodiments, the molar ratio of the combinedmolar amount of the first compounds to the combined molar amount of thesecond compounds is about 1:10, about 1:9, about 1:8, about 1:7, about1:6, about 1:5, about 1:4, about 1:3, about 1:2, or about 1:1. Incertain embodiments, when the molar ratio of the combined molar amountof the first compounds (e.g., amines) to the combined molar amount ofthe second compounds (e.g., esters) is greater than 1, the stickiness ofthe resulting retinal patch is increased.

In some embodiments, the first compound comprising more than onenucleophilic group and the therapeutic agent are pre-mixed in an aqueousbuffer. Once pre-mixing is complete, the second compound comprising morethan one electrophilic group is added to the pre-mixture. Shortly afterfinal mixing, the hydrogel polymer is delivered to the target site. Incertain embodiments, the optional radiopaque material is added to thepre-mix, the second compound, or to the mixture just before delivery ofthe hydrogel polymer mixture to the target site.

In other embodiments, the second compound comprising more than oneelectrophilic group and the therapeutic agent are pre-mixed in anaqueous buffer. Once pre-mixing is complete, the first compoundcomprising more than one nucleophilic group is added to the pre-mixture.Shortly after final mixing, the hydrogel polymer is delivered to thetarget site. In certain embodiments, the optional radiopaque material isadded to the pre-mix, the first compound, or to the mixture just beforedelivery of the hydrogel polymer mixture to the target site.

In some embodiments, the first compound comprising more than onenucleophilic group and the second compound comprising more than oneelectrophilic group are mixed together in an aqueous buffer in the pHrange of about 5.0 to about 9.5, whereby a biocompatible hydrogelpolymer is formed. In certain embodiments, the first compound comprisingmore than one nucleophilic group and/or the second compound comprisingmore than one electrophilic group are individually diluted in an aqueousbuffer in the pH range of about 5.0 to about 9.5, wherein the individualdilutions or neat monomers are mixed, whereby a biocompatible hydrogelpolymer is formed. In some embodiments, the aqueous buffer is in the pHrange of about 6.0 to about 8.5. In certain embodiments, the aqueousbuffer is in the pH range of about 8.

In certain embodiments, the concentration of the monomers in the aqueousis from about 1% to about 100%. In some embodiments, the dilution isused to adjust the viscosity of the monomer dilution. In certainembodiments, the concentration of the monomers in the aqueous buffer isabout 1%, is about 1.5%, is about 2%, is about 2.5%, is about 3%, isabout 3.5%, is about 4%, is about 4.5%, about 5%, about 6%, about 7%,about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about95%, or about 100%.

In some embodiments, the electrophilic and nucleophilic monomers aremixed in such ratio that there is a slight excess of electrophilicgroups present in the mixture. In certain embodiments, this excess isabout 10%, about 5%, about 2%, about 1%, about 0.9%, about 0.8%, about0.7%, about 0.6%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, about0.1%, or less than 0.1%.

In certain embodiments, the gelling time or curing time of thebiocompatible hydrogel polymer is controlled by the selection of thefirst and second compounds. In some embodiments, the concentration ofnucleophilic or electrophilic groups in the first or second compoundinfluences the gelling time of the in vivo gelling ophthalmicpre-formulation. In certain embodiments, temperature influences thegelling time of the in vivo gelling ophthalmic pre-formulation. In someembodiments, the type of aqueous buffer influences the gelling time ofthe in vivo gelling ophthalmic pre-formulation. In certain embodiments,the concentration of the aqueous buffer influences the gelling time ofthe in vivo gelling ophthalmic pre-formulation. In some embodiments, thenucleophilicity and/or electrophilicity of the nucleophilic andelectrophilic groups of the monomers influences the gelling time of thein vivo gelling ophthalmic pre-formulation.

In some embodiments, the gelling time or curing time of thebiocompatible hydrogel polymer is controlled by the pH of the aqueousbuffer. In certain embodiments, the gelling time decreases with anincrease in pH. In some embodiments, the gelling time decreases with anincrease in buffer concentration. In certain embodiments, the gellingtime decreases with an increase in temperature. In some embodiments, thegelling time decreases with an increase in solution (monomer)concentration.

In certain embodiments, the gelling time is between about 20 seconds and10 minutes. In some embodiments, the gelling time is less than 30minutes, less than 20 minutes, less than 10 minutes, less than 5minutes, less than 4.8 minutes, less than 4.6 minutes, less than 4.4minutes, less than 4.2 minutes, less than 4.0 minutes, less than 3.8minutes, less than 3.6 minutes, less than 3.4 minutes, less than 3.2minutes, less than 3.0 minutes, less than 2.8 minutes, less than 2.6minutes, less than 2.4 minutes, less than 2.2 minutes, less than 2.0minutes, less than 1.8 minutes, less than 1.6 minutes, less than 1.4minutes, less than 1.2 minutes, less than 1.0 minutes, less than 0.8minutes, less than 0.6 minutes, or less than 0.4 minutes. In certainembodiments, the pH of the aqueous buffer is from about 5 to about 9.5.In some embodiments, the pH of the aqueous buffer is from about 7.0 toabout 9.5. In specific embodiments, the pH of the aqueous buffer isabout 8. In some embodiments, the pH of the aqueous buffer is about 5,about 5.5, about 6.0, about 6.5, about 6.6, about 6.7, about 6.8, about6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5,about 7.6, about 7.8, about 7.9, about 8.0, about 8.1 about 8.2 about8.3, about 8.4, about 8.5, about 9.0, or about 9.5.

In certain embodiments, the gelling time or curing time of thebiocompatible hydrogel polymer is controlled by the type of aqueousbuffer. In some embodiments, the aqueous buffer is a physiologicallyacceptable buffer. In certain embodiments, aqueous buffers include, butare not limited to, aqueous saline solutions, phosphate buffered saline,borate buffered saline, a combination of borate and phosphate bufferswherein each component is dissolved in separate buffers,N-2-Hydroxyethylpiperazine-N′-2-hydroxypropanesulfonic acid (HEPES),3-(N-Morpholino) propanesulfonic acid (MOPS),2-([2-Hydroxy-1,1-bis(hydroxymethyl)ethyl]amino)ethanesulfonic acid(TES),3-[N-tris(Hydroxy-methyl)ethylamino]-2-hydroxyethyl]-1-piperazinepropanesulfonicacid (EPPS), Tris[hydroxymethyl]-aminomethane (THAM), andTris[hydroxymethyl]methyl aminomethane (TRIS). In some embodiments, thethiol-ester chemistry (e.g., ETTMP nucleophile with SGA or SGelectrophile) is performed in borate buffer. In certain embodiments, theamine-ester chemistry (NH2 or AA nucleophile with SGA or SGelectrophile) is performed in phosphate buffer.

In some embodiments, the tack of the retinal patch is about 40 mN. Incertain embodiments, the tack of the retinal patch is between about 20mN and about 100 mN. In some embodiments, the tack of the retinal patchis between about 30 mN and about 50 mN. In certain embodiments, thefirmness of the retinal patch is between about 30 g to about 100 g. Insome embodiments, the firmness of the retinal patch is between about 45g to about 90 g. In certain embodiments, the elastic modulus of theretinal patch is between about 50 Pa to about 500 Pa. In someembodiments, the elastic modulus of the retinal patch is between about100 Pa to about 400 Pa.

In certain embodiments, the first compound and the second compound donot react with the therapeutic agent during formation of thebiocompatible hydrogel polymer. In some embodiments, the therapeuticagent remains unchanged after polymerization of the first and secondcompounds (i.e., monomers). In certain embodiments, the therapeuticagent does not change the properties of the hydrogel polymer. In someembodiments, the physiochemical properties of the therapeutic agent andthe hydrogel polymer formulation are not affected by the polymerizationof the monomers.

Area of for Treatment—Target Sites

In certain embodiments, the target site is inside a mammal. In someembodiments, the target site is inside a human being. In certainembodiments, the target site is on the human body. In some embodiments,the target site is accessible through surgery. In certain embodiments,the target site is accessible through minimally invasive surgery. Insome embodiments, the target site is accessible through an endoscopicdevice. In certain embodiments, the target site is in or on an eye. Insome embodiments, a method of treating a retinal tear, hole, or retinaldetachment comprises delivering the in vivo gelling ophthalmicpre-formulation to the site of the hole, tear, or detachment under localanesthesia. In certain embodiments, the in vivo gelling ophthalmicpre-formulation is delivered through a sharp 24 to 28 gauge needle.

In some embodiments, an in vivo gelling ophthalmic pre-formulation or abiocompatible hydrogel polymer is used as a sealant or adhesive with orwithout a therapeutic agent. In certain embodiments, the in vivo gellingophthalmic pre-formulation or biocompatible hydrogel polymer is used toseal retinal tears inside a human eye. In other embodiments, the in vivogelling ophthalmic pre-formulation or biocompatible hydrogel polymer isused to fill cavities in the human body, e.g., an eye as partial orcomplete replacement of the vitreous humour.

Delivery of the Hydrogel Formulation to a Target Site

In some embodiments, the in vivo gelling ophthalmic pre-formulation isdelivered as an in vivo gelling ophthalmic pre-formulation to a targetsite through a catheter or a needle to form a biocompatible hydrogelpolymer at the target site. In certain embodiments, the needle orcatheter is attached or part of a delivery device.

In other embodiments, the in vivo gelling ophthalmic pre-formulation isdelivered to the target site in the eye using a syringe and needle. Insome embodiments, a delivery device is used to deliver the in vivogelling ophthalmic pre-formulation to the target site. In someembodiments, the needle has an outer diameter of about 4 mm, about 3.8mm, about 3.6 mm, about 3.4 mm, about 3.2 mm, about 3.0 mm, about 2.8mm, about 2.6 mm, about 2.4 mm, about 2.2 mm, about 2.0 mm, about 1.8mm, about 1.6 mm, about 1.4 mm, about 1.2 mm, about 1.0 mm, about 0.8mm, or about 0.6 mm. In preferred embodiments, the needle has an outerdiameter of about 1.2 mm or less. In certain embodiments, the viscosityof the in vivo gelling ophthalmic pre-formulation is close to theviscosity of water when delivering the mixture to the site of the tumorthrough the catheter. In some embodiments, the in vivo gellingophthalmic pre-formulation forming the biocompatible hydrogel furthercomprises a pharmaceutically acceptable viscosity enhancer to ensurethat the pre-formulation stays in place at the target site during thegelling process.

In certain embodiments, between 1 and 3 mL of the in vivo gellingophthalmic pre-formulation optionally comprising a therapeutic agent isdelivered to a target site. In some embodiments, about 12 mL, about 11mL, about 10 mL, about 9 mL, about 8 mL, about 7.5 mL, about 7.0 mL,about 6.5 mL, about 6.0 mL, about 5.5 mL, about 5.0 mL, about 4.5 mL,about 4.0 mL, about 3.5 mL, about 3.0 mL, about 2.5 mL, about 2.0 mL,about 1.5 mL, about 1.0 mL, about 0.5 mL, about 0.2 mL, about 0.1 mL,about 0.05 mL or about 0.01 mL in vivo gelling ophthalmicpre-formulation optionally comprising a therapeutic agent is deliveredto a target site. In certain embodiments, less than 12 mL, less than 11mL, less than 10 mL, less than 9 mL, less than 8 mL, less than 7.5 mL,less than 7.0 mL, less than 6.5 mL, less than 6.0 mL, less than 5.5 mL,less than 5.0 mL, less than 4.5 mL, less than 4.0 mL, less than 3.5 mL,less than 3.0 mL, less than 2.5 mL, less than 2.0 mL, less than 1.5 mL,less than 1.0 mL, less than 0.5 mL, less than 0.2 mL, less than 0.1 mL,less than 0.05 mL, or less than 0.01 mL in vivo gelling ophthalmicpre-formulation optionally comprising a therapeutic agent is deliveredto a target site. In certain embodiments, about 0.05 to 5 mL in vivogelling ophthalmic pre-formulation optionally comprising a therapeuticagent is delivered to a target site.

In some embodiments, the gelling time of the biocompatible hydrogelpolymer is set according to the preference of the doctor delivering thehydrogel polymer mixture to a target site. In most instances, aphysician delivers the hydrogel polymer mixture to the target within 15to 30 seconds. In some embodiments, the hydrogel polymer mixture gelsafter delivery at the target site, covering the target site.

In some embodiments, the gelling time or curing time of thebiocompatible hydrogel polymer is controlled by the pH of the aqueousbuffer. In certain embodiments, the gelling time is between about 20seconds and 10 minutes. In preferred embodiments, the gelling time isabout 90 seconds. In some embodiments, the gelling time is less than 120minutes, less than 90 minutes, less than 60 minutes, less than 50minutes, less than 40 minutes, less than 30 minutes, less than 20minutes, less than 10 minutes, less than 9 minutes, less than 8 minutes,less than 7 minutes, less than 6 minutes, less than 5 minutes, less than4.8 minutes, less than 4.6 minutes, less than 4.4 minutes, less than 4.2minutes, less than 4.0 minutes, less than 3.8 minutes, less than 3.6minutes, less than 3.4 minutes, less than 3.2 minutes, less than 3.0minutes, less than 2.8 minutes, less than 2.6 minutes, less than 2.4minutes, less than 2.2 minutes, less than 2.0 minutes, less than 1.8minutes, less than 1.6 minutes, less than 1.5 minutes, less than 1.4minutes, less than 1.2 minutes, less than 1.0 minutes, less than 0.8minutes, less than 0.6 minutes, or less than 0.4 minutes. In certainembodiments, the gelling time is more than 120 minutes, more than 90minutes, more than 60 minutes, more than 50 minutes, more than 40minutes, more than 30 minutes, more than 20 minutes, more than 10minutes, more than 9 minutes, more than 8 minutes, more than 7 minutes,more than 6 minutes, more than 5 minutes, more than 4.8 minutes, morethan 4.6 minutes, more than 4.4 minutes, more than 4.2 minutes, morethan 4.0 minutes, more than 3.8 minutes, more than 3.6 minutes, morethan 3.4 minutes, more than 3.2 minutes, more than 3.0 minutes, morethan 2.8 minutes, more than 2.6 minutes, more than 2.4 minutes, morethan 2.2 minutes, more than 2.0 minutes, more than 1.8 minutes, morethan 1.6 minutes, more than 1.5 minutes, more than 1.4 minutes, morethan 1.2 minutes, more than 1.0 minutes, more than 0.8 minutes, morethan 0.6 minutes, or more than 0.4 minutes. In some embodiments, thegelling time is about 120 minutes, about 90 minutes, about 60 minutes,about 50 minutes, about 40 minutes, about 30 minutes, about 20 minutes,about 10 minutes, about 9 minutes, about 8 minutes, about 7 minutes,about 6 minutes, about 5 minutes, about 4.8 minutes, about 4.6 minutes,about 4.4 minutes, about 4.2 minutes, about 4.0 minutes, about 3.8minutes, about 3.6 minutes, about 3.4 minutes, about 3.2 minutes, about3.0 minutes, about 2.8 minutes, about 2.6 minutes, about 2.4 minutes,about 2.2 minutes, about 2.0 minutes, about 1.8 minutes, about 1.6minutes, about 1.5 minutes, about 1.4 minutes, about 1.2 minutes, about1.0 minutes, about 0.8 minutes, about 0.6 minutes, or about 0.4 minutes.

In certain embodiments, the pH of the aqueous buffer is from about 5.0to about 9.5. In some embodiments, the pH of the aqueous buffer is fromabout 6.0 to about 8.5. In specific embodiments, the pH of the aqueousbuffer is about 8.0. In some embodiments, the pH is about 5, about 5.1,about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about5.8, about 5.9, about 6, about 6.1, about 6.2, about 6.3, about 6.4,about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7, about7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7,about 7.8, about 7.9, about 8, about 8.1, about 8.2, about 8.3, about8.4, about 8.5, about 8.6, about 8.7, about 8.9, about 9, about 9.1about 9.2, about 9.3, about 9.4, or about 9.5.

In certain embodiments, the gelling time or curing time of thebiocompatible hydrogel polymer is controlled by the selection of thefirst and second compounds. In some embodiments, the concentration ofnucleophilic or electrophilic groups in the first or second compoundinfluences the gelling time of the in vivo gelling ophthalmicpre-formulation.

In some embodiments, curing of the biocompatible hydrogel polymer isverified post-administration. In certain embodiments, the verificationis performed in vivo at the delivery site. In other embodiments, theverification is performed ex vivo. In some embodiments, curing of thebiocompatible hydrogel polymer is verified visually. A lack of flow ofthe biocompatible hydrogel polymer indicates that the biocompatiblehydrogel polymer has gelled and the hydrogel is sufficiently cured. Infurther embodiments, curing of the biocompatible hydrogel polymer isverified by evaluation of the residue in the delivery device, forinstance the residue in the catheter of the bronchoscope or otherendoscopic device, or the residue in the syringe used to deliver thebiocompatible hydrogel polymer. In other embodiments, curing of thebiocompatible hydrogel polymer is verified by depositing a small sample(e.g., ˜1 mL) on a piece of paper or in a small vessel and subsequentevaluation of the flow characteristics after the gelling time haspassed.

In some embodiments, the in vivo gelling ophthalmic pre-formulationoptionally comprising one or more therapeutic agents is delivered to thetarget site so that the pre-formulation mostly covers the target site.In certain embodiments, the in vivo gelling ophthalmic pre-formulationsubstantially covers an exposed portion of diseased tissue. In someembodiments, the in vivo gelling ophthalmic pre-formulation does notspread to any other location intentionally. In some embodiments, the invivo gelling ophthalmic pre-formulation substantially covers diseasedtissue and does not significantly cover healthy tissue. In certainembodiments, the biocompatible hydrogel polymer does not significantlycover healthy tissue. In some embodiments, in vivo gelling ophthalmicpre-formulation gels over the target site and thoroughly covers diseasedtissue. In some embodiments, the biocompatible hydrogel polymer adheresto tissue.

Bioabsorbance of the Hydrogel

In some embodiments, the biocompatible hydrogel polymer is abioabsorbable polymer. In certain embodiments, the biocompatiblehydrogel polymer is bioabsorbed within about 5 to 30 days. In someembodiments, the biocompatible hydrogel polymer is bioabsorbed withinabout 30 to 180 days. In preferred embodiments, the biocompatiblehydrogel polymer is bioabsorbed within about 1 to 70 days. In someembodiments the biocompatible hydrogel polymer is bioabsorbed withinabout 365 days, 180 days, about 150 days, about 120 days, about 90 days,about 80 days, about 70 days, about 60 days, about 50 days, about 40days, about 35 days, about 30 days, about 28 days, about 21 days, about14 days, about 10 days, about 7 days, about 6 days, about 5 days, about4 days, about 3 days, about 2 days, or about 1 day. In certainembodiments the biocompatible hydrogel polymer is bioabsorbed withinless than 365 days, 180 days, less than 150 days, less than 120 days,less than 90 days, less than 80 days, less than 70 days, less than 60days, less than 50 days, less than 40 days, less than 35 days, less than30 days, less than 28 days, less than 21 days, less than 14 days, lessthan 10 days, less than 7 days, less than 6 days, less than 5 days, lessthan 4 days, less than 3 days, less than 2 days, or less than 1 day. Insome embodiments the biocompatible hydrogel polymer is bioabsorbedwithin more than 365 days, 180 days, more than 150 days, more than 120days, more than 90 days, more than 80 days, more than 70 days, more than60 days, more than 50 days, more than 40 days, more than 35 days, morethan 30 days, more than 28 days, more than 21 days, more than 14 days,more than 10 days, more than 7 days, more than 6 days, more than 5 days,more than 4 days, more than 3 days, more than 2 days, or more than 1day. In some embodiments, the biocompatible hydrogel polymer issubstantially non-bioabsorbable.

The biocompatible hydrogel polymer is slowly bioabsorbed, dissolved, andor excreted. In some instances, the rate of bioabsorption is controlledby the number of ester groups in the biocompatible and/or biodegradablehydrogel polymer. In other instances, the higher the concentration ofester units is in the biocompatible hydrogel polymer, the longer is itslifetime in the body. In further instances, the electron density at thecarbonyl of the ester unit controls the lifetime of the hydrogel polymerin the body. In certain instances, biocompatible hydrogel polymerswithout ester groups are essentially not biodegradable. In additionalinstances, the molecular weight of the first and second compoundscontrols the lifetime of the hydrogel polymer in the body. In furtherinstances, the number of ester groups per gram of polymer controls thelifetime of the hydrogel polymer in the body.

In some instances, the lifetime of the hydrogel polymer can be estimatedusing a model, which controls the temperature and pH at physiologicallevels while exposing the hydrogel polymer to a buffer solution. Incertain instances, the biodegradation of the hydrogel polymer issubstantially non-enzymatic degradation.

In some embodiments, the selection of reaction conditions determines thedegradation time of the hydrogel polymer. In certain embodiments, theconcentration of the first compound and second compound monomersdetermines the degradation time of the resulting hydrogel polymer. Insome instances, a higher monomer concentration leads to a higher degreeof cross-linking in the resulting hydrogel polymer. In certaininstances, more cross-linking leads to a later degradation of thehydrogel polymer.

In certain embodiments, the composition of the linker in the firstand/or second compound influences the speed of degradation of theresulting hydrogel polymer. In some embodiments, the more ester groupsare present in the hydrogel polymer, the faster the degradation of thehydrogel polymer. In certain embodiments, the higher the concentrationof mercaptopropionate (ETTMP), acetate amine (AA), glutarate orsuccinate (SG or SS) monomers, the faster the rate of degradation.

Retinal Patch or Suture in the Treatment of Retinal Disease

In some embodiments, the in vivo gelling ophthalmic pre-formulationdescribed herein is delivered to a target site of an eye to treatretinal detachment. In certain embodiments, the in vivo gellingophthalmic pre-formulation is delivered to a target site of an eye totreat blindness. In some embodiments, the in vivo gelling ophthalmicpre-formulation forms a biocompatible retinal patch at the target siteinside the eye. In certain embodiments, the in vivo gelling ophthalmicpre-formulation acts as a retinal glue at a target site inside the eye.In some embodiments, the in vivo gelling ophthalmic pre-formulationforms a retinal suture. In certain embodiments, the retinal patch,retinal glue, or retinal suture gels at least in part at the target siteinside the eye. In some embodiments, the retinal patch, retinal glue, orretinal suture gels at least in part at a retinal tear inside the eye.In certain embodiments, the retinal patch, retinal glue, or retinalsuture polymerizes at least in part at the target site inside the eye.In some embodiments, the retinal patch, retinal glue, or retinal suturepolymerizes at least in part at a retinal tear inside the eye. In someembodiments, the retinal patch, retinal glue, or retinal suture adheresat least partially to the target site.

In certain embodiments, the in vivo gelling hydrogel polymer is used asa “liquid suture” or as a drug delivery platform to transportmedications directly to the targeted site in the eye. In someembodiments, the spreadability, viscosity, optical clarity, and adhesiveproperties of the hydrogel formulation are optimized to create materialsideal as liquid sutures for the treatment of retinal detachment(re-attachment of detached retina). In certain embodiments, the gel timeis controlled from 50 seconds to 15 minutes.

Control of Release Rate of a Therapeutic Agent

In some embodiments, the biocompatible hydrogel polymer slowly deliversa therapeutic agent to a target site by diffusion and/or osmosis overtime ranging from hours to days. In certain embodiments, the drug isdelivered directly to the target site. In some embodiments, theprocedure of delivering a biocompatible hydrogel polymer comprising atherapeutic agent to a target site is repeated several times, if needed.In other embodiments, the therapeutic agent is released from thebiocompatible hydrogel polymer through biodegradation of the hydrogelpolymer. In some embodiments, the therapeutic agent is released througha combination of diffusion, osmosis, and/or hydrogel degradationmechanisms. In certain embodiments, the release profile of thetherapeutic agent from the hydrogel polymer is unimodal. In someembodiments, the release profile of the therapeutic agent from thehydrogel polymer is bimodal. In certain embodiments, the release profileof the therapeutic agent from the hydrogel polymer is multimodal.

In some embodiments, the therapeutic agent is released from thebiocompatible hydrogel polymer though diffusion or osmosis. In certainembodiments, the therapeutic agent is substantially released from thebiocompatible hydrogel polymer within 180 days. In some embodiments, thetherapeutic agent is substantially released from the biocompatiblehydrogel polymer within 14 days. In certain embodiments, the therapeuticagent is substantially released from the biocompatible hydrogel polymerwithin 24 hours. In some embodiments, the therapeutic agent issubstantially released from the biocompatible hydrogel polymer withinone hour. In certain embodiments, the therapeutic agent is substantiallyreleased from the biocompatible hydrogel polymer within about 180 days,about 150 days, about 120 days, about 90 days, about 80 days, about 70days, about 60 days, about 50 days, about 40 days, about 35 days, about30 days, about 28 days, about 21 days, about 14 days, about 10 days,about 7 days, about 6 days, about 5 days, about 4 days, about 3 days,about 2 days, about 1 day, about 0.5 day, about 6 hours, about 4 hours,about 2 hours, about or 1 hour. In some embodiments, the therapeuticagent is substantially released from the biocompatible hydrogel polymerwithin more than 180 days, more than 150 days, more than 120 days, morethan 90 days, more than 80 days, more than 70 days, more than 60 days,more than 50 days, more than 40 days, more than 35 days, more than 30days, more than 28 days, more than 21 days, more than 14 days, more than10 days, more than 7 days, more than 6 days, more than 5 days, more than4 days, more than 3 days, more than 2 days, more than 1 day, more than0.5 day, more than 6 hours, more than 4 hours, more than 2 hours, morethan or 1 hour. In certain embodiments, the therapeutic agent issubstantially released from the biocompatible hydrogel polymer withinless than 180 days, less than 150 days, less than 120 days, less than 90days, less than 80 days, less than 70 days, less than 60 days, less than50 days, less than 40 days, less than 35 days, less than 30 days, lessthan 28 days, less than 21 days, less than 14 days, less than 10 days,less than 7 days, less than 6 days, less than 5 days, less than 4 days,less than 3 days, less than 2 days, less than 1 day, less than 0.5 day,less than 6 hours, less than 4 hours, less than 2 hours, less than or 1hour. In some embodiments, the therapeutic agent is substantiallyreleased from the biocompatible hydrogel polymer within about one day toabout fourteen days. In certain embodiments, the therapeutic agent issubstantially released from the biocompatible hydrogel polymer withinabout one day to about 70 days.

In some embodiments, the therapeutic agent is a biomolecule and therelease of the biomolecule from the hydrogel polymer is controlled bythe composition of the hydrogel polymer. In certain embodiments, thebiomolecule is released when the hydrogel polymer starts to degrade. Insome embodiments, the pore size of the hydrogel polymer is small enoughto prevent the early phase release of the biomolecule (i.e., releasebefore the degradation of the hydrogel polymer). In certain embodiments,the pore size of the hydrogel polymer is large enough to allow the earlyphase release of the biomolecule. In some embodiments, the ratio of thepore size of the hydrogel polymer to the size of the biomoleculedetermines the release rate of the biomolecule.

Exemplary Antifungals

In some embodiments, the biocompatible hydrogel polymer comprises anantifungal agent as the therapeutic agent. In certain embodiments, theantifungal agent is a polyene antifungal, an imidazole, triazole, orthiazole antifungal, a triazole antifungal, a thiazole antifungal, anallylamine derivative, or an echinocandin derivative. Examples ofantifungal agents include, but are not limited to, Polyene derivativeslike natamycin, rimocidin, filipin, nystatin, amphotericin B, candicin,hamycin; Imidazole derivatives like miconazole, ketoconazole,clotrimazole, econazole, omoconazole, bifonazole, butoconazole,fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole,tioconazole; Tetrazole derivatives like fluconazole, itraconazole,isavuconazole, posaconazole, voriconzaole, terconazole, albaconazole;Thiazole derivatives like abafungin; Allylamine derivative liketerbifine, naftifine, butenafine; Echinocandin derivatives likeanidulafungin, caspofungin, micafungin; Other antifungals likepolygodial, benzoic acid, ciclopirox, tonaftate, undecylenic acid,flycytosine, griseofulvin, haloprogin, sodium bicarbonate, pirctoneolamine, zinc pyrithione, selenium sulfide, tar, or tea tree oil.

Exemplary Antibiotics

In some embodiments, the biocompatible hydrogel polymer comprises anantibiotic. In certain embodiments, the antibiotic agent is aaminoglycoside, ansamycin, carbacephem, carbapenem, cephalosporin,glycopeptide, lincosamide, lipopeptide, macrolide, monobactam,nitrofurans, penicillin, polypeptide, quinolone, sulfonamide, ortetracycline. Examples of antibiotic agents include, but are not limitedto, Aminoglycoside derivatives like amikacin, gentamicin, kanamycin,neomycin, netilmicin, tobramicin, paromomycin; Ansamycin derivativeslike geldanamycin, herbimycin; Carbacephem derivatives like loracarbef,Carbapenem derivatives like ertapenem, doripenem, imipenem, meropenem;Cephalosporin derivatives like cefadroxil, cefazolin, cefalotin,cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime,cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime,ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime,ceftobiprole; Glycopeptide derivatives like teicoplanin, vancomycin,telavancin; Lincosamides like clindamycin, lincomycin; Lipopeptidederivatives like daptomycin; Macrolide derivatives like azithromycin,clarithromycin, dirithromycin, erythromycin, roxithromycin,troleandomycin; telithreomycin, spectinomycin; Monobactam derivativeslike aztreonam; Nitrofuran derivatives like furazolidone,nitrofurantoin; Penicillin derivatives like amoxicillin, ampicillin,azlocillin, carbinicillin, cloxacillin, dicloxacillin, flucloxacillin,mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillinV, piperacillin, temocillin, ticarcillin; Penicillin combinations likeamoxicillin/clavulanate, ampicillin/sulbactam, piperacillin/tazobactam,ticarcillin/clavulanate; Polypeptide derivatives like bacitracin,colistin, polymyxin B; Quinolone derivatives like ciprofloxacin,enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin,nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin,sparfloxacin, temafloxacin; Sulfonamide derivatives like mafenide,sulfonamidochrysoidine, sulfacetamide, sulfadiazine, silversulfadiazine, sulfamethoxazole, sulfanilimide, sulfasalazine,sulfisoxazole, trimethoprim, trimethoprim/sulfamethoxazole; Tetracyclinderivatives like demeclocycline, doxycycline, minocycline,oxytetracycline, tetracycline; Derivatives against mycobacteria likeclofazimine, dapsone, capreomycin, cycloserine, ethambutol, ethioamide,isoniazid, pyrazinamide, rifampin, refampicin, rifabutin, rifapentine,streptomycin; or other antibiotic agents like arsphenamine,chloramphenicol, fosfomycin, fusidic acid, linezolid, metronidazole,mupirocin, platensimycin, quinupristin/dalfopristin, rifaximin,thiampheniol, tigecycline, tinidazole.

Exemplary Antiviral Agents

In some embodiments, the biocompatible hydrogel polymer comprises anantiviral agent. In certain embodiments, the antiviral agent is anucleoside reverse transcriptase inhibitor, a non-nucleoside reversetranscriptase inhibitor, a fusion inhibitor, an integrase inhibitor, anucleoside analog, a protease inhibitor, a reverse transcriptaseinhibitor. Examples of antiviral agents include, but are not limited to,abacavir, aciclovir, acyclovir, adefovir, amantadine, amprenavir,ampligen, arbidol, atazanavir, boceprevir, cidofovir, darunavir,delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine,enfuvirtide, entecavir, famciclovir, fomivirsen, fosamprenavir,foscarnet, fosfonet, ganciclovir, ibacitabine, imunovir, idoxuridine,imiquimod, indinavir, inosine, interferon type III, interferon type II,interferon type I, interferon, lamivudine, lopinavir, loviride,maraviroc, moroxydine, methisazone, nelfinavir, nevirapine, nexavir,oseltamivir, peginterferon alfa-2a, penciclovir, peramivir, pleconaril,podophyllotoxin, raltegravir, ribavirin, rimantadine, ritonavir,pyramidine, saquinavir, stavudine, tea tree oil, tenofovir, tenofovirdisoproxil, tipranavir, trifluridine, trizivir, tromantadine, truvada,valaciclovir (Valtrex), valganciclovir, vicriviroc, vidarabine,viramidine, zalcitabine, zanamivir, zidovudine.

Exemplary Immunosuppressive Agents

In some embodiments, the biocompatible hydrogel polymer comprises animmunosuppressive agent. In certain embodiments, the immunosuppressiveagent is a calcinuerin inhibitor, mTor inhibitor, an anti-proliferativeagent (e.g., an alkylating agent or an antimetabolite), aglucocorticosteroid, an antibody, or an agent acting on immunophilins.Examples of immunosuppressive agents include, but are not limited to,Calcineurin inhibitors like ciclosporin, tacrolimus; mTOR inhibitorslike sirolimus, everolimus; Anti-proliferatives like azathioprine,mycophenolic acid; Corticosteroids like prednisolone, hydrocortisone;Monoclonal anti-IL-2Rα receptor antibodies like basiliximab, daclizumab;Polyclonal anti-T-cell antibodies like antithymocyte globulin (ATG),anti-lymphocyte globulin (ALG); Monoclonal anti-CD20 antibodies likerituximab; Interleukin inhibitors like daclizumab, basiliximab,anakinra, rilonacept, ustekinumab, mepolizumab, tocilizumab,canakinumab, briakinumab; Tumor necrosis factor alpha (TNF-α) inhibitorslike etanercept, infliximab, afelimomab, adalimumab, certolizumab pegol,golimumab; Selective immunosuppressants like muromonab-CD3,antilymphocyte immunoglobulin (horse), antithymocyte immunoglobulin(rabbit), mycophenolic acid, sirolimus, leflunomide, alefacept,everolimus, gusperimus, efalizumab, abetimus, natalizumab, abatacept,eculizumab, belimumab, fingolimod, belatacept; or Otherimmunosuppressants like azathioprine, thalidomide, methotrexate,lenalidomide

Exemplary Hemostasis Agents

In some embodiments, the biocompatible hydrogel polymer comprises ahemostasis agent (or antihemorrhagic agent). In certain embodiments, thehemostasis agent is an antifibrinolytic (amino acid or proteinaseinhibitor), a vitamin K, fibrinogen, a local hemostatic, or a bloodcoagulation factor. Examples of hemostasis agents include, but are notlimited to, Amino acids like aminocaproic acid, tranexamic acid,aminomethylbenzoic acid; Proteinase inhibitors like aprotinin, alfa1antitrypsin, C1-inhibitor, camostat; Vitamin K like phytomenadione,menadione; Fibrinogen like Human fibrinogen; Local hemostatics likeabsorbable gelatin sponge, oxidized cellulose, tetragalacturonic acidhydroxymethylester, adrenalone, thrombin, collagen, calcium alginate,epinephrine, human fibrinogen; Blood coagulation factors likecoagulation factor IX, II, VII and X in combination, coagulation factorVIII, factor VIII inhibitor bypassing activity, coagulation factor IX,coagulation factor VII, von Willebrand factor and coagulation factorVIII in combination, coagulation factor XIII, eptacog alfa, nonacogalfa, thrombin; Other systemic hemostatics like etamsylate,carbazochrome, batroxobin, romiplostim, eltrombopag.

Exemplary Non-Steroidal Anti-Inflammatory Agents

In some embodiments, the biocompatible hydrogel polymer comprises ananti-inflammatory agent. In certain embodiments, the anti-inflammatoryagent is a non-steroidal anti-inflammatory agent. In other embodiments,the anti-inflammatory agent is a glucocorticosteroid. In someembodiments, the non-steroidal anti-inflammatory agent is abutylpyrazolidine, an acetic acid derivative, oxicam, propionic acidderivative, fenamate, or coxib. Examples of anti-inflammatory agentsinclude, but are not limited to, Butylpyrazolidines like phenylbutazone,mofebutazone, oxyphenbutazone, clofezone, kebuzone; Acetic acidderivatives and related substances like indometacin, sulindac, tolmetin,zomepirac, diclofenac, alclofenac, bumadizone, etodolac, lonazolac,fentiazac, acemetacin, difenpiramide, oxametacin, proglumetacin,ketorolac, aceclofenac, bufexamac, indometacin combinations, diclofenaccombinations; Oxicams like piroxicam, tenoxicam, droxicam, lornoxicam,meloxicam; Propionic acid derivatives like ibuprofen, naproxen,ketoprofen, fenoprofen, fenbufen, benoxaprofen, suprofen, pirprofen,flurbiprofen, indoprofen, tioprofenoic acid, oxaprozin, ibuproxam,dexibuprofen, flunoxaprofen, alminoprofen, dexketoprofen, naproxcinod;Fenamates like mefenamic acid, tolfenamic acid, flufenamic acid,meclofenamic acid; Coxibs like celecoxib, rofecoxib, valdecoxib,parecoxib, etoricoxib, lumiracoxib; Other antiinflammatory andantirheumatic agents like nabumetone, niflumic acid, azapropazone,glucosamine, benzydamine, glucosaminoglycan polysulfate, proquazone,orgotein, nimesulide, feprazone, diacerein, morniflumate, tenidap,oxaceprol, chondroitin sulfate.

Exemplary Analgesics and Anesthetics

In some embodiments, the biocompatible hydrogel polymer comprises ananalgesic or anesthetic agent. In certain embodiments, the analgesic oranesthetic agent comprises paracetamol, an opiate, diproqualone,phenazone, cocaine, or lidocaine. In certain embodiments, the opioid isa natural opium alkaloid, phenylpiperidine derivative,diphenylpropylamine derivative, benzomorphan derivative, oripavinderivative, or morphinan derivative. In some embodiments, the analgesicis a salicylic acid derivative, pyrazolone, or anilide. In otherembodiments, the analgesic is an ergot alkaloid, corticosteroidderivative, or selective serotonin (5HT1) agonist. Examples of localanesthetics include, but are not limited to, Esters of aminobenzoic acidlike metabutethamine, procaine, tetracaine, chloroprocaine, benzocaine;Amides like bupivacaine, lidocaine, mepivacaine, prilocaine,butanilicaine, cinchocaine, etidocaine, articaine, ropivacaine,levobupivacaine, tetracaine, chloroprocaine, benzocaine; Esters ofbenzoic acid like cocaine; Other local anesthetics like ethyl chloride,dyclonine, phenol, capsaicin.

Exemplary Proteins and Other Biomolecules

In some embodiments, the biocompatible hydrogel polymer comprises aprotein or other biomolecule. Examples of proteins and otherbiomolecules include, but are not limited to abarelix, abatacept,acarbose, adalimumab, alglucosidase alfa, Antihemophilic FactorRecombinant, antithrombin recombinant lyophilized powder forreconstitution, belatacept, belimumab, bevacizumab, botulinum toxin typeA, canakinumab, certolizumab pegol, Cetrotide, cetuximab, chorionichuman recombinant gonadotropin, coagulation Factor IX (recombinant),collagenase clostridium histolyticum, conjugated estrogens,Cyanocobalamin, darbepoetin alfa, denosumab, Diphtheria and TetanusToxoids and Acellular Pertussis Vaccine Adsorbed, Diptheria and TetanusToxoids and Acellular Pertussis Vaccine Absorbed, dornase alfa,drotrecogin alfa [activated]), ecallantide, eculizumab, enfuvirtide,enoxaparin sodium, epoetin alfa, etanercept, exenatide, filgrastim,follitropin alfa, follitropin beta, Fragmin, galsulfase, gemtuzumabozogamicin, glatiramer acetate, Glucagon, golimumab, goserelin acetate,Haemophilus b Conjugate Vaccine—Tetanus Toxoid Conjugate, histrelinacetate, ibritumomab tiuxetan, idursulfase, incobotulinumtoxin A,infliximab, Influenza Virus Vaccine, insulin derivatives, insulinaspart, insulin glargine [rDNA origin], insulin lispro, interferonalfacon-1, interferon beta-1a, Interferon beta-1b, ipilimumab, JapaneseEncephalitis Vaccine—Inactivated—Adsorbed, lanreotide acetate,laronidase, leuprolide acetate for depot suspension, leuprolide acetate,linagliptin, liraglutide, mecasermin, menotropins, methoxy polyethyleneglycol-epoetin beta, natalizumab, ofatumumab, omalizumab,onabotulinumtoxin A, palivizumab, pancrelipase, pancrelipase,panitumumab, pegaptanib, pegfilgrastim, peginterferon alfa-2a,peginterferon alfa-2b, pegloticase, pegvisomant, pentosan polysulfatesodium, pramlintide, quadrivalent human papillomavirus (types 6, 11, 16,18) recombinant vaccine, ranibizumab, rasburicase, Recombinant HumanPapillomavirus Bivalent (Types 16 and 18) Vaccine, recombinantInterferon alfa-2b, reteplase, Rituximab, romiplostim, sargramostim,secretin, sevelamer carbonate, sevelamer hydrochloride, sipuleucel-T,somatropin, somatropin [rDNA origin], teriparatide, tocilizumab,trastuzumab, triptorelin pamoate, ustekinumab, velaglucerase alfa forinjection.

In certain embodiments, the biocompatible hydrogel polymer comprises aprotein as a pharmaceutically active biomolecule. Examples of proteinsinclude, but are not limited to, octreotide, eptifibatide, desmopressin,leuprolide/leuprorelin, goserelin, ciclosporin, bivalirudin, glucagon,calcitonin, teriparatide, enfuvirtide, ecallantide, romiplostim. In someembodiments, the biocompatible polymer comprises a recombinant proteinas a pharmaceutically active biomolecule. Examples of recombinantproteins include, but are not limited to, insulin, lepirudin,somatropin, aldesleukin, interferon gamma 1b, anakinra, interferon alpha2b, interferon beta 1b, interferon beta 1a, PEG interferon alpha 2a,filgrastim, pegfilgrastim, oprelvekin, reteplase, denileukin diftitox,follitropin alfa, recFSH, thyrotropin alfa, imiglucerase, becaplermin,sargramostim, darbepoetin, erythropoietin, DNAse, Factor VIIa, FactorIX, Factor XIII, drotrecogin, alteplase, tenecteplase, moroctocog alfa(BDDrFVIII), Factor VIII-2, Factor VIII, peginteferon, ribavarin,clostridial collagenese, alglucosidase alpha2, incobotulinumtoxina,pegloticase, palifermin, galsulfase, idursulfase. In certainembodiments, the biocompatible hydrogel polymer comprises an antibody asa pharmaceutically active biomolecule. Examples of antibodies include,but are not limited to, etanercept, abciximab, gemtuzumab, rituximab,adalimumab, palivizumab, trastuzumab, bevacizumab, natalizumab,omalizumab, infliximab, alemtuzumab, efalizumab, cetuximab, golimumab,abobotulinumtoxina, canakinumab, ustekinumab, ofatumumab, certolizumabpegol, tocilizumab, denosumab, abatacept, ranibizumab, panitumumab,eculizumab, brentixumab, iplimumab, belimumab, rilonacept.

EXAMPLES

The following specific examples are to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever.

The following general characteristics of the monomers and polymers areneeded to be successful for bonding the retina without causing anyadverse effects.

Monomers Property Characteristics 1 In vivo Could be polymerized insidemammalian polymerizable eyes 2 Reaction mixture Physiological to 8.0 pHrange pH 3 Reaction tempera- Ambient to body temperature ture 4Formulation Two or three component system; Mixed physical formimmediately prior to use 5 Mixing time for Few seconds (~10 sec) thereaction to start 6 Gel formation Gel formation time ranges from 10 timeseconds to 120 seconds 7 Solution Solution viscosity ranges from 1 toviscosity 800 cps 8 Sterilization ETO to E-beam sterilizable capability9 Localized Ideal for localized delivery for small delivery molecules,large molecules and cells Option- Stability of All small molecule drugsand proteins ally drugs in studied so far, have been found to beformulation stable mixture

Below are some adhesive polymer characteristics.

Adhesive Property Characteristics 1 Tissue adhesion Sticky formulations,physicochemical characteristics ideal for bonding to retina 2 OpticalClarity Optically clear materials 3 Polymer hardness Similar to retinaand the surrounding tissues 4 Bioabsorption Time About 2 weeks (up to 10years for drug delivery) 5 Biocompatibility Highly biocompatible; passedall the subjected ISO 10993 tests 6 Polymer Non-cytotoxic formulationscytotoxicity Option- Small molecule Small drug molecules elution can beally elution controlled and thus pharmaceutical drugs could also bedelivered using the formulations, if needed Option- Compatibility withHighly compatible due to physiological ally proteins and Cells pH of thepolymers

For applications on-site, desired gel times are under 120 seconds.Additionally, the viscosity should be high enough to prevent excessivespreading around the target treatment area, but low enough to enter anysmall cavities at the site. Furthermore, the reaction buffers should beclose to physiological conditions. The desired degradation time andpolymer pore size will vary based on the application. The polymer shouldbe elastic and strong enough to resist fragmentation in the body.

The chemical components of the polymers are listed in Table 1. Thechemical monomers will be referred to by their abbreviations. SeveralUSP grade ophthalmic use approved viscosity enhancing agents werepurchased from Sigma-Aldrich and were stored at 25° C. They includemethylcellulose (Methocel® MC, 10-25 MPA·S) abbreviated as MC;hypromellose (hydroxypropylmethylcellulose 2910) abbreviated as HPMC;and povidone K-30 (polyvinylpyrrolidone) abbreviated as PVP. Themonomers were stored at 5° C. and allowed to warm to room temperaturebefore use, which typically took 30 minutes. After use the contents werepurged with N₂ for approximately 30 seconds before sealing with parafilmand returning to 5° C.

A 0.15 M phosphate buffer was made by dissolving 9.00 g (0.075 mol)NaH₂PO₄ in 500 mL of distilled water at 25° C. with magnetic stirring.The pH was then adjusted to 7.99 with the dropwise addition of 50%aqueous NaOH. Several other phosphate buffers were prepared in a similarfashion: 0.10 M phosphate at pH 9, 0.10 M phosphate at pH 7.80, 0.10 Mphosphate at 7.72, 0.10 M phosphate at pH 7.46, 0.15 M phosphate at pH7.94, 0.15 M phosphate at pH 7.90, 0.4 M phosphate at pH 9, and 0.05 Mphosphate at pH 7.40.

A sterile 0.10 M phosphate buffer at pH 7.58 with 0.30% HPMC wasprepared for use in kits. First, 1.417 g HPMC was dissolved in 471 mL of0.10 M phosphate buffer at pH 7.58 by vigorous shaking. The viscoussolution was allowed to clarify overnight. The solution was filteredthrough a 0.22 μm filter (Corning #431097) with application of lightvacuum. The viscosity of the resulting solution was measured to be 8.48cSt+/−0.06 at 20° C.

Phosphate buffered saline (PBS) was prepared by dissolving two PBStablets (Sigma Chemical, P4417) in 400 mL of distilled water at 25° C.with vigorous shaking. The solution has the following composition andpH: 0.01 M phosphate, 0.0027 M potassium chloride, 0.137 M sodiumchloride, pH 7.46.

A 0.058 M phosphate buffer was made by dissolving 3.45 g (0.029 mol) ofNaH₂PO₄ in 500 mL of distilled water at 25° C. with magnetic stirring.The pH was then adjusted to 7.97 with the dropwise addition of 50%aqueous NaOH. A 0.05 M borate buffer was made by dissolving 9.53 g(0.025 mol) of Na₂B₄O₇.10 H2O in 500 mL of distilled water at 25° C.with magnetic stirring. The pH was then adjusted to 7.93 or 8.35 withthe dropwise addition of 6.0 N HCl.

The amine or thiol component (typically in the range of 0.1 mmol armsequivalents) was added to a 50 mL centrifuge tube. A volume of reactionbuffer was added to the tube via a pipette such that the finalconcentration of solids in solution was about 5 percent. The mixture wasgently swirled to dissolve the solids before adding the appropriateamount of ester or epoxide. Immediately after adding the ester orepoxide, the entire solution was shaken for 10 seconds before letting itrest.

The gel time for all cases was measured starting from the addition ofthe ester or epoxide until the gelation of the solution. The gel pointwas noted by pipetting 1 mL of the reaction mixture and observing thedropwise increase in viscosity. Degradation of the polymers wasperformed by the addition of 5 to 10 mL of phosphate buffered saline toca. 5 g of the material in a 50 mL centrifuge tube and incubating themixture at 37° C. The degradation time was measured starting from theday of addition of the phosphate buffer to complete dissolution of thepolymer into solution.

TABLE 1 Components used in formulations. Components Technical NameETTMP-1300 Ethoxylated trimethylolpropane tri(3- mercaptopropionate)4ARM-5k-SH 4ARM PEG Thiol (pentaerythritol) 4ARM-2k-NH2 4ARM PEG Amine(pentaerythritol), HCl Salt, MW 2000 4ARM-5k-NH2 4ARM PEG Amine(pentaerythritol), HCl Salt, MW 5000 8ARM-20k-NH2 8ARM PEG Amine(hexaglycerol), HCl Salt, MW 20000 4ARM-20k-AA 4ARM PEG Acetate AmineHCl Salt, MW 20000 8ARM-20k-AA 8ARM PEG Acetate Amine (hexaglycerol) HClSalt, MW 20000 8ARM-20k-AA 8ARM PEG Acetate Amine (hexaglycerol) TFASalt, MW 20000 4ARM-10k-SG 4ARM PEG Succinimidyl Glutarate(pentaerythritol), MW 10000 8ARM-15k-SG 8ARM PEG Succinimidyl Glutarate(hexaglycerol), MW 15000 4ARM-20k-SGA 4ARM PEG Succinimidyl Glutaramide(pentaerythritol), MW 20000 4ARM-10k-SS 4ARM PEG Succinimidyl Succinate(pentaerythritol), MW 10000 EJ-190 Sorbitol polyglycidyl ether MC MethylCellulose (Methocel ® MC) HPMC Hypromellose(Hydroxypropylmethylcellulose) PVP Povidone (polyvinylpyrrolidone)

Example 1 Manufacture of Hydrogel (Amine-Ester Chemistry)

A solution of 8ARM-20K-NH2 was prepared in a Falcon tube by dissolvingabout 0.13 g solid monomer in about 2.5 mL of sodium phosphate buffer(buffer pH 7.36). The mixture was shaken for about 10 seconds at ambienttemperature until complete dissolution was obtained. The Falcon tube wasallowed to stand at ambient temperature. In another Falcon tube, 0.10 gof 8ARM-15K-SG was dissolved in the same phosphate buffer as above. Themixture was shaken for about 10 seconds and at this point all the powderdissolved. The 8ARM-15K-SG solution was poured immediately into the8ARM-20K-NH2 solution and a timer was started. The mixture was shakenand mixed for about 10 seconds and a 1 mL solution of the mixture waspipetted out using a mechanical high precision pipette. The gel time of1 mL liquid was collected and then verified with the lack of flow forthe remaining liquids. The gel time data of the formulation was recordedand was about 90 seconds.

Example 2 Manufacture of Hydrogel (Amine-Ester Chemistry)

A solution of amines was prepared in a Falcon tube by dissolving about0.4 g solid 4ARM-20k-AA and about 0.2 g solid 8ARM-20k-NH2 in about 18mL of sodium phosphate buffer (buffer pH 7.36). The mixture was shakenfor about 10 seconds at ambient temperature until complete dissolutionwas obtained. The Falcon tube was allowed to stand at ambienttemperature. To this solution, 0.3 g of 8ARM-15K-SG was added. Themixture was shaken to mix for about 10 seconds until all the powderdissolved. 1 mL of the mixture was pipetted out using a mechanical highprecision pipette. The gel time of the formulation was collected usingthe process described above. The gel time was about 90 seconds.

Example 3 Manufacture of Hydrogel (Thiol-Ester Chemistry

A solution of ETTMP-1300 was prepared in a Falcon tube by dissolvingabout 0.04 g monomer in about 5 mL of sodium borate buffer (buffer pH8.35). The mixture was shaken for about 10 seconds at ambienttemperature until complete dissolution was obtained. The Falcon tube wasallowed to stand at ambient temperature. To this solution, 0.20 g of8ARM-15K-SG was added. The mixture was shaken for about 10 seconds untilthe powder dissolved. 1 mL of the mixture was pipetted out using amechanical high precision pipette. The gel time was found to be about 70seconds.

Example 4 Manufacture of Hydrogel (Thiol-Epoxide Chemistry)

A solution of ETTMP-1300 was prepared in a Falcon tube by dissolvingabout 0.04 g monomer in about 5 mL of sodium borate buffer (buffer pH8.35). The mixture was shaken for about 10 seconds at ambienttemperature until complete dissolution was obtained. The Falcon tube wasallowed to stand at ambient temperature. To this solution, 0.10 g ofEJ-190 was added. The mixture was shaken for about 10 seconds untilcomplete dissolution is obtained. 1 mL of the mixture was pipetted outusing a mechanical high precision pipette. The gel time was found to beabout 6 minutes.

Example 5 In Vitro Bioabsorbance Testing

A 0.10 molar buffer solution of pH 7.40 was prepared with deionizedwater. A 50 mL portion of this solution was transferred to a Falcontube. A sample polymer was prepared in a 20 cc syringe. After curing, a2-4 mm thick slice was cut from the polymer slug and was placed in theFalcon tube. A circulating water bath was prepared and maintained at 37°C. The Falcon tube with polymer was placed inside the water bath andtime was started. The dissolution of the polymer was monitored andrecorded. The dissolution time ranged from 1-90 days depending on thetype of sample polymer.

Example 6 Gelling and Degradation Times of Amine-Ester Polymers

Amines studied were 8ARM-20k-NH2 and 4ARM-5k-NH2. The formulationdetails and material properties are given in Table 2. With 8ARM-20k-NH2,it was found that a phosphate buffer with 0.058 M phosphate and pH of7.97 was necessary to obtain acceptable gel times of around 100 seconds.Using a 0.05 M phosphate buffer with a pH of 7.41 resulted in a morethan two-fold increase in gel time (270 seconds).

With the 8ARM-20k-NH2, the ratio of 4ARM-10k-SS to 4ARM-20k-SGA wasvaried from 50:50 to 90:10. The gel time remained consistent, but therewas a marked shift in degradation time around a ratio of 80:20. Forformulations with ratios of 75:25 and 50:50, degradation times spiked toone month and beyond. Using lower amounts of 4ARM-20k-SGA (80:20, 85:15,90:10) resulted in degradation times of less than 7 days.

As a comparison, the 4ARM-5k-NH2 was used in a formulation with a ratioof 4ARM-10k-SS to 4ARM-20k-SGA of 80:20. As was expected, thedegradation time remained consistent, which suggests that the mechanismof degradation was unaffected by the change in amine. However, the geltime increased by 60 seconds, which may reflect the relativeaccessibility of reactive groups in a high molecular weight 8ARM amineand a low molecular weight 4ARM amine.

TABLE 2 Gel and degradation times for varying 4ARM-10k- SS/4ARM-20k-SGAratios with 8ARM-15k-SG ester. Phosphate Reaction Degrada- Ratio ofBuffer Con- Gel tion 4ARM-10k-SS/ centration Time Time Components4ARM-20k-SGA and pH (s) (days) 8ARM-20k-NH2 50/50 0.05M 270 N/A4ARM-10k-SS, pH 7.41 4ARM-20k-SGA 8ARM-20k-NH2 50/50 0.058M 100 >414ARM-10k-SS, pH 7.97 4ARM-20k-SGA 8ARM-20k-NH2 75/25 0.058M 90 294ARM-10k-SS, pH 7.97 4ARM-20k-SGA 8ARM-20k-NH2 80/20 0.058M 100 74ARM-10k-SS, pH 7.97 4ARM-20k-SGA 4ARM-5k-NH2 80/20 0.058M 160 64ARM-10k-SS, pH 7.97 4ARM-20k-SGA 8ARM-20k-NH2 85/15 0.058M 100 54ARM-10k-SS, pH 7.97 4ARM-20k-SGA 8ARM-20k-NH2 90/10 0.058M 90 64ARM-10k-SS, pH 7.97 4ARM-20k-SGA

Example 7 Gelling and Degradation Times of Thiol-Ester Polymers

Thiols studied were 4ARM-5k-SH and ETTMP-1300. The formulation detailsand material properties are given in Table 3. It was found that a 0.05 Mborate buffer with a pH of 7.93 produced gel times of around 120seconds. Increasing the amount of 4ARM-20k-SGA in the formulationincreased the gel time to 190 seconds (25:75 ratio of 4ARM-10k-SS to4ARM-20k-SGA) up to 390 seconds (0:100 ratio of 4ARM-10k-SS to4ARM-20k-SGA). Using a 0.05 M borate buffer with a pH of 8.35 resultedin a gel time of 65 seconds, about a two-fold decrease in gel time.Thus, the gel time may be tailored by simply adjusting the pH of thereaction buffer.

The ratio of 4ARM-10k-SS to 4ARM-20k-SGA was varied from 0:100 to 100:0.In all cases, the degradation time did not vary significantly and wastypically between 3 and 5 days. It is likely that degradation isoccurring via alternate pathways.

TABLE 3 Gel and degradation times for varying 4ARM-10k-SS/4ARM- 20k-SGAratios with 4ARM-5k-SH and ETTMP-1300 thiols. Phosphate ReactionDegrada- Ratio of Buffer Con- Gel tion 4ARM-10k-SS/ centration Time TimeComponents 4ARM-20k-SGA and pH (s) (days) 4ARM-5k-SH 50/50 0.05M 65 N/A4ARM-10k-SS, pH 8.35 4ARM-20k-SGA 4ARM-5k-SH 50/50 0.05M 120 44ARM-10k-SS, pH 7.93 4ARM-20k-SGA 4ARM-5k-SH 75/25 0.05M 125 44ARM-10k-SS, pH 7.93 4ARM-20k-SGA 4ARM-5k-SH 90/10 0.05M 115 44ARM-10k-SS, pH 7.93 4ARM-20k-SGA 4ARM-5k-SH 25/75 0.05M 190 44ARM-10k-SS, pH 7.93 4ARM-20k-SGA 4ARM-5k-SH 10/90 0.05M 200 44ARM-10k-SS, pH 7.93 4ARM-20k-SGA ETTMP-1300  0/100 0.05M 390 34ARM-20k-SGA 4ARM-5k-SH 100/0  0.05M 120 4 4ARM-10k-SS pH 7.93

Example 8 Gelling and Degradation Times of Amine-Ester and Thiol-EsterPolymers

An amine (4ARM-5k-NH2) and a thiol (4ARM-5k-SH) were studied with theester 4ARM-10k-SG. The formulation details and material properties aregiven in Table 4. A 0.058 M phosphate buffer with a pH of 7.97 yielded agel time of 150 seconds with the amine. A 0.05 M borate buffer with a pHof 8.35 produced a gel time of 75 seconds with the thiol.

The amine-based polymer appeared to show no signs of degradation, as wasexpected from the lack of degradable groups. However, the thiol-basedpolymer degraded in 5 days. This suggests that degradation is occurringthrough alternate pathways, as was observed in the thiol formulationswith 4ARM-10k-SS and 4ARM-20k-SGA (vida supra).

TABLE 4 Gel and degradation times for amines and thiols with 4ARM-10k-SGformulations. Gel Reaction Buffer Type, Time Degradation ComponentsConcentration, and pH (s) Time (days) 4ARM-5k-NH2 & Phosphate (0.058M,pH 7.97) 150 In- 4ARM-10k-SG definite 4ARM-5k-SH & Borate (0.05M, pH8.35) 75 5 4ARM-10k-SG

Example 9 Gelling and Degradation Times of Thiol-Sorbitol PolyglycidylEther Polymers

With ETTMP-1300 conditions such as high pH (10), high solutionconcentration (50%), or high borate concentration (0.16 M) werenecessary for the mixture to gel. Gel times ranged from around 30minutes to many hours. The conditions that were explored include: pHfrom 7 to 12; solution concentration from 5% to 50%; borateconcentration from 0.05 M to 0.16 M; and thiol to epoxide ratios from1:2 to 2:1.

The high pH necessary for the reaction to occur could result indegradation of the thiol. Thus, a polymer with EJ-190 and 4ARM-5k-SH wasprepared. A 13% solution formulation exhibited a gel time of 230 secondsat a pH of between 9 and 10. The degradation time was 32 days. At alower pH of around 8, the mixture exhibited gel times in the range of 1to 2 hours.

Example 10 General Procedure for the Preparation of In VivoPolymerizable Materials

Several representative sticky formulations are listed in Table 5 alongwith specific reaction details for the preparation of in vivopolymerizable materials. The polymers were prepared by first dissolvingthe amine component in phosphate buffer or the thiol component in boratebuffer. The appropriate amount of the ester component was then added andthe entire solution was mixed vigorously for 10 to 20 seconds. The geltime was measured starting from the addition of the ester until thegelation of the solution.

TABLE 5 (A) Summary of the reaction details for several representativesticky formulations without viscosity enhancer; (B) more detailedtabulation of a selection of the reaction details including moles(degradation times were measured in phosphate buffered saline (PBS) at37° C.). (A) Amine or Thiol/ Ester Degradation Molar % Gel Time TimeComponents Ratio Buffer Solution (s) (days) 8ARM-20k-NH2 3 0.15Mphosphate, 3 130 N/A 4ARM-20K-SGA pH 7.99 8ARM-20k-NH2 1/3 0.15Mphosphate, 3 300 N/A 4ARM-20K-SGA pH 7.99 8ARM-20k-NH2 3 0.15Mphosphate, 8 50 N/A 4ARM-10K-SS pH 7.99 8ARM-20k-NH2 1/3 0.15Mphosphate, 8 80 N/A 4ARM-10K-SS pH 7.99 4ARM-20K-AA/ 3 0.15M phosphate,5 210 1 to 3 8ARM-20k-NH2 pH 7.99 (75/25) 4ARM-20K-SGA 4ARM-20K-AA/ 50.15 M phosphate, 10 180 1 to 3 8ARM-20k-NH2 pH 7.99 (75/25)4ARM-20K-SGA 4ARM-5K-NH2 5 0.10M phosphate, 10 160  7 4ARM-10K-SG pH7.80 4ARM-5K-NH2 5 0.10M phosphate, 20 160 1 to 3 4ARM-10K-SS pH 7.804ARM-5K-NH2 3 0.10M phosphate, 5 160 13 4ARM-10K-SG pH 7.80 4ARM-5K-NH25 0.15M phosphate, 20 80 7 4ARM-10K-SG pH 7.99 4ARM-5K-NH2 5 0.15Mphosphate, 30 70 10 4ARM-10K-SG pH 7.99 4ARM-5K-NH2 5 0.15M phosphate,19 60 53 4ARM-20K-SGA pH 7.99 4ARM-5K-NH2 5 0.15M phosphate, 12 70 534ARM-20K-SGA pH 7.99 4ARM-5K-NH2 1/5 0.15M phosphate, 19 160 154ARM-10K-SG pH 7.99 4ARM-SH-5K 5 0.05M borate, 20 120 2 to 4 4ARM-10K-SGpH 7.93 4ARM-NH2-2K 5 0.10M phosphate, 10 120 15 8ARM-15K-SG pH 7.464ARM-NH2-2K 7 0.10M phosphate, 30 150 N/A 4ARM-20K-SGA pH 7.80 (B)Polymer % Wt Arms Solution Components MW Mmoles (g) Arm mmoles Eq (w/v)8ARM-20k-NH2 20000 1000 0.075 8 0.00375 0.03 4ARM-20k-SGA 20000 10000.05 4 0.0025 0.01 Buffer Volume (phosphate) 4.1 3.0 8ARM-20k-NH2 200001000 0.025 8 0.00125 0.01 4ARM-20k-SGA 20000 1000 0.15 4 0.0075 0.03Buffer Volume (phosphate) 5.8 3.0 8ARM-20k-NH2 20000 1000 0.3 8 0.0150.12 4ARM-10k-SS 10000 1000 0.1 4 0.01 0.04 Buffer Volume (phosphate) 58.0 8ARM-20k-NH2 20000 1000 0.1 8 0.005 0.04 4ARM-10k-SS 10000 1000 0.34 0.03 0.12 Buffer Volume (phosphate) 5 8.0

TABLE 6 Gel times for the 8ARM-20k-NH2/4ARM-20k-SGA(1/1) sticky polymersincluding HPMC as viscosity enhancer with varying buffers andconcentrations. Amine/Ester % Gel Time Components Molar Ratio BufferSolution (min) 8ARM-20k-NH2 1 0.10M 4.8 1.5 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.80 8ARM-20k-NH2 1 0.10M 4.8 3.5 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.46 8ARM-20k-NH2 1 0.05M 4.8 4.5 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.42 8ARM-20k-NH2 1 0.05M 4 5.5 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.42 8ARM-20k-NH2 1 0.05M 3 8.5 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.42 8ARM-20k-NH2 1 0.05M 4.8 6.75 4ARM-20K-SGA phosphate, 0.3%HPMC pH 7.24 8ARM-20k-NH2 1 0.05M 3 12 4ARM-20K-SGA phosphate, 0.3% HPMCpH 7.24 8ARM-20k-NH2 1 0.05M 2.5 15.5 4ARM-20K-SGA phosphate, 0.3% HPMCpH 7.24

Gel times ranged from 60 to 300 seconds and were found to be easilytuned by adjusting the reaction buffer pH, buffer concentration, orpolymer concentration. An example of gel time control for a singleformulation is shown in Table 6, where the gel time for the8ARM-20k-NH2/4ARM-20k-SGA (1/1) polymer was varied from 1.5 to 15.5minutes.

In some instances, the stickiness of the polymers originates from amismatching in the molar equivalents of the components. A variety ofsticky materials using combinations of 4 or 8 armed amines of molecularweights between 2 and 20 thousand and 4 or 8 armed esters of molecularweights between 10 and 20 thousand were created. It was found that incomparison with the 8 armed esters, the 4 armed esters resulted instickier materials. For the amine component, it was found that smallermolecular weights led to stickier materials and higher amine to estermolar ratios.

A mismatch (amine to ester molar ratio) of at least 3 was required toqualitatively sense stickiness. More preferably, a ratio of around 5produced a desirable level of stickiness combined with polymer strength.Polymers with amine to ester molar ratios higher than 5 may be formed aswell, but some reaction conditions, such as the polymer concentration,may need to be adjusted to obtain a reasonable gel time. Furthermore, itwas found that the use of a viscosity enhanced solution improves thepolymers by increasing their strength and elasticity, allowing forhigher amine to ester molar ratios (Example 11, Table 8).

The materials formed were typically transparent and elastic. Stickinesswas tested for qualitatively by touch. Thus, a sticky material adheredto a human finger or other surface and remained in place until removed.Degradation times varied from 1 to 53 days. In certain instances, thepolymer properties, such as gel and degradation times, pore sizes,swelling, etc. may be optimized for different applications withoutlosing the stickiness.

Example 11 General Procedure for the Preparation of Solutions withEnhanced Viscosity

Polymer solutions with enhanced viscosities were prepared by theaddition of a viscosity enhancing agent to the reaction buffer. Table 8Blists the viscosity enhancing agents studied, including observations onthe properties of the formed polymers. Stock solutions of reactionbuffers were prepared with varying concentrations of methylcellulose(MC), hypromellose (HPMC) or polyvinylpyrrolidone (PVP). As an example,a 2% (w/w) HPMC solution in buffer was made by adding 0.2 g of HPMC to9.8 mL of 0.10 M phosphate buffer at pH 7.80, followed by vigorousshaking. The solution was allowed to stand overnight. Buffer solutionswith HPMC concentrations ranging from 0.01% to 2.0% were prepared in asimilar fashion. Buffer solutions with PVP concentrations ranging from5% to 20% and buffer solutions with MC concentrations ranging from 1.0to 2.0% were also prepared by a similar method.

The polymers were formed in the same method as described above in thegeneral procedures for the preparation of the sticky materials (Example10). A typical procedure involved first dissolving the amine componentin the phosphate buffer containing the desired concentration ofviscosity enhancing agent. The appropriate amount of the ester componentwas then added and the entire solution was mixed vigorously for 10 to 20seconds. The gel time was measured starting from the addition of theester until the gelation of the solution.

Several representative formulations are listed in Table 7 along withspecific reaction details. The percent of degradable acetate aminecomponent by mole equivalents is represented by a ratio designated inparenthesis. For example, a formulation with 75% degradable amine willbe written as 8ARM-20k-AA/8ARM-20k-NH2 (75/25). The polymer was preparedby first dissolving the amine component in phosphate buffer. Theappropriate amount of the ester component was then added and the entiresolution was mixed vigorously for 10 to 20 seconds. The gel time wasmeasured starting from the addition of the ester until the gelation ofthe solution.

The gel time is dependent on several factors: pH, buffer concentration,polymer concentration, temperature and the monomers used. Previousexperiments have shown that the extent of mixing has little effect onthe gel time once the components are in solution, which typically takesup to 10 seconds. FIG. 1 shows the effect of monomer addition on bufferpH. For the 8ARM-20k-NH2 & 4ARM-20k-SGA formulation, the buffer pH dropsslightly from 7.42 to 7.36 upon addition of the monomers. For the8ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA formulation, the bufferpH drops from 7.4 to 7.29 upon addition of the monomers. The additionaldecrease in the pH was found to originate from acidic residues in thedegradable acetate amine. The same pH drop phenomenon was observed forthe 4ARM-20k-AA amine. In certain instances, a quality controlspecification on the acetate amine solution pH may be required toimprove the consistency of degradable formulations.

FIG. 2 depicts the effect of reaction buffer pH on gel times. The geltimes increase with an increase in the concentration of hydronium ionsin an approximately linear fashion. More generally, the gel timesdecrease with an increase in the buffer pH. FIG. 3 shows the effect ofreaction buffer phosphate concentration on gel times. The gel timesdecrease with an increase in the phosphate concentration. FIG. 4illustrates the effect of polymer concentration on gel times. The geltimes decrease significantly with an increase in the polymerconcentration. At low polymer concentrations where the gel time isgreater than 5 minutes, hydrolysis reactions of the ester begin tocompete with the formation of the polymer. The effect of temperature ongel times appears to follow the Arrhenius equation, as seen in FIG. 5.The gel time is directly related to the extent of reaction of thepolymer solution and so this behavior is not unusual.

In FIG. 6, the rheology of the polymers during the gelation process isshown as a function of the percent time to the gel point. Thus, 100%represents the gel point and 50% represents half the time before the gelpoint. The viscosity of the reacting solution remains relativelyconstant until about 80% of the gel point. After that point, theviscosity increases dramatically, representing the formation of thesolid gel.

FIG. 7 shows the gel time stability of a single formulation using thesame lot of monomers over the course of about a year. The monomers werehandled according to the standard protocol outlined above. The gel timesremained relatively stable; some variations in the reaction buffer mayaccount for differences in the gel times.

TABLE 7 (A) Summary of the reaction details for several representativesticky formulations; (B) more detailed tabulation of a selection of thereaction details including moles (degradation times were measured inphosphate buffered saline (PBS) at 37° C.). (A) % Degradation ComponentsBuffer Solution Gel Time (s) Time (days) 4ARM-20k-AA/8ARM-20k-NH2 0.10Mphosphate, 5 150 21 (60/40) pH 7.80 4ARM-20k-SGA4ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 5 150 21 (60/40) pH 7.804ARM-20k-SGA 0.3% HPMC 8ARM-20k-NH2 0.10M phosphate, 4.8 100 N/A4ARM-20k-SGA pH 7.80 0.3% HPMC 8ARM-20k-NH2 0.10M phosphate, 4.8 70 488ARM-15k-SG pH 7.80 0.3% HPMC 4ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate,4.8 110 12 (60/40) pH 7.80 8ARM-15k-SG 0.3% HPMC4ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 20 160 21 (60/40) pH 7.804ARM-20k-SGA 0.3% HPMC 8ARM-20k-NH2 0.10M phosphate, 4.8 90 N/A4ARM-20k-SGA pH 7.80 8ARM-20k-NH2 0.10M phosphate, 4.8 80 N/A4ARM-20k-SGA pH 7.80 1.0% HPMC 8ARM-20k-NH2 0.10M phosphate, 4.8 210 N/A4ARM-20k-SGA pH 7.46 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 4.8 270 N/A4ARM-20k-SGA pH 7.42 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 4 330 N/A4ARM-20k-SGA pH 7.42 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 3 510 N/A4ARM-20k-SGA pH 7.42 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 4.8 405 N/A4ARM-20k-SGA pH 7.24 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 3 720 N/A4ARM-20k-SGA pH 7.24 0.3% HPMC 8ARM-20k-NH2 0.05M phosphate, 2.5 930 N/A4ARM-20k-SGA pH 7.24 0.3% HPMC 8ARM-20k-AA 0.10M phosphate, 4.8 90 64ARM-20k-SGA pH 7.46 HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10Mphosphate, 4.8 100 16 (75/25) pH 7.46 4ARM-20k-SGA HPMC (0.3%)8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8 95 256 (60/40) pH 7.46(estimated) 4ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10Mphosphate, 4.8 120 N/A (50/50) pH 7.46 4ARM-20k-SGA HPMC (0.3%)8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8 100 21 (70/30) pH 7.464ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8100 28 (65/35) pH 7.46 4ARM-20k-SGA HPMC (0.3%) 8ARM-20k-NH2 0.10Mphosphate, 4.8 90 N/A 4ARM-20k-SGA pH 7.80 1.5% HPMC8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8 90 16 (75/25) pH 7.464ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8105 21 (70/30) pH 7.46 4ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH20.10M phosphate, 4.8 120 N/A (50/50) pH 7.46 4ARM-20k-SGA HPMC (0.3%)8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8 70 7 (70/30) pH 7.468ARM-15k-SG HPMC (0.3%) 4ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8260 10 (70/30) pH 7.46 4ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH20.10M phosphate, 4.8 70 17 (60/40) pH 7.46 8ARM-15k-SG HPMC (0.3%)8ARM-20k-AA 0.10M phosphate, 4.8 85 7 4ARM-20k-SGA pH 7.46 HPMC (0.3%)8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.8 95 13 (70/30) pH 7.464ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 4.895 10 (75/25) pH 7.46 4ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH20.10M phosphate, 4 110 10 (75/25) pH 7.58 4ARM-20k-SGA HPMC (0.3%)8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 3.5 150 9 (75/25) pH 7.584ARM-20k-SGA HPMC (0.3%) 8ARM-20k-AA/8ARM-20k-NH2 0.10M phosphate, 3 1908 (75/25) pH 7.58 4ARM-20k-SGA HPMC (0.3%) (B) Polymer % Wt ArmsSolution Components MW Mmoles (g) Arm mmoles Eq (w/v) 8ARM-20k-NH2 200001000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 1000 0.08 4 0.004 0.016Buffer Volume (phosphate) 2.5 4.8 Viscosity Enhancer 0.3% HPMC8ARM-20k-NH2 20000 1000 0.08 8 0.004 0.032 8ARM-15k-SG 15000 1000 0.06 80.004 0.032 Buffer Volume (phosphate) 2.9 4.8 Viscosity Enhancer 0.3%HPMC 8ARM-20k-AA 20000 1000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 10000.08 4 0.004 0.016 Buffer Volume (phosphate) 2.5 4.8 Viscosity Enhancer0.3% HPMC 4ARM-20k-AA 20000 1000 0.06 4 0.003 0.012 8ARM-20k-NH2 200001000 0.02 8 0.001 0.008 4ARM-20k-SGA 20000 1000 0.1 4 0.005 0.02 BufferVolume (phosphate) 3.6 5.0 Viscosity Enhancer 0.3% HPMC 4ARM-20k-AA20000 1000 0.12 4 0.006 0.024 8ARM-20k-NH2 20000 1000 0.04 8 0.002 0.0168ARM-15k-SG 15000 1000 0.075 4 0.005 0.02 Buffer Volume (phosphate) 4.94.8 Viscosity Enhancer 0.3% HPMC 8ARM-20k-AA 20000 1000 0.06 8 0.0030.024 8ARM-20k-NH2 20000 1000 0.02 8 0.001 0.008 4ARM-20k-SGA 20000 10000.16 4 0.008 0.032 Buffer Volume (phosphate) 5 4.8 Viscosity Enhancer0.3% HPMC 8ARM-20k-AA 20000 1000 0.03 8 0.0015 0.012 8ARM-20k-NH2 200001000 0.02 8 0.001 0.008 4ARM-20k-SGA 20000 1000 0.1 4 0.005 0.02 BufferVolume (phosphate) 3.1 4.8 Viscosity Enhancer 0.3% HPMC 8ARM-20k-AA20000 1000 0.02 8 0.001 0.008 8ARM-20k-NH2 20000 1000 0.02 8 0.001 0.0084ARM-20k-SGA 20000 1000 0.08 4 0.004 0.016 Buffer Volume (phosphate) 2.54.8 Viscosity Enhancer 0.3% HPMC 8ARM-20k-AA 20000 1000 0.035 8 0.001750.014 8ARM-20k-NH2 20000 1000 0.015 8 0.00075 0.006 4ARM-20k-SGA 200001000 0.1 4 0.005 0.02 Buffer Volume (phosphate) 3.1 4.8 ViscosityEnhancer 0.3% HPMC 8ARM-20k-AA 20000 1000 0.039 8 0.00195 0.01568ARM-20k-NH2 20000 1000 0.021 8 0.00105 0.0084 4ARM-20k-SGA 20000 10000.12 4 0.006 0.024 Buffer Volume (phosphate) 3.75 4.8 Viscosity Enhancer0.3% HPMC 8ARM-20k-AA 20000 1000 0.09 8 0.0045 0.036 8ARM-20k-NH2 200001000 0.03 8 0.0015 0.012 4ARM-20k-SGA 20000 1000 0.24 4 0.012 0.048Buffer Volume (phosphate) 9 4.0 Viscosity Enhancer 0.3% HPMC 8ARM-20k-AA20000 1000 0.075 8 0.00375 0.03 8ARM-20k-NH2 20000 1000 0.025 8 0.001250.01 4ARM-20k-SGA 20000 1000 0.2 4 0.01 0.04 Buffer Volume (phosphate)8.55 3.5 Viscosity Enhancer 0.3% HPMC 8ARM-20k-AA 20000 1000 0.06 80.003 0.024 8ARM-20k-NH2 20000 1000 0.02 8 0.001 0.008 4ARM-20k-SGA20000 1000 0.16 4 0.008 0.032 Buffer Volume (phosphate) 8 3.0 ViscosityEnhancer 0.3% HPMCCytotoxicity & Hemolysis Evaluation

Several polymer samples were sent out to NAMSA for cytotoxicity andhemolysis evaluation. Cytotoxic effects were evaluated according to ISO10993-5 guidelines. Hemolysis was evaluated according to proceduresbased on ASTM F756 and ISO 10993-4.

The polymer 8ARM-20k-NH2 & 4ARM-20k-SGA at 4.8% solution with 0.3% HPMCwas found to be non-cytotoxic and non-hemolytic. The polymer8ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA at 4.8% solution with0.3% HPMC was found to be non-cytotoxic and non-hemolytic. In addition,formulations involving 4ARM-20kAA and 8ARM-15k-SG were alsonon-cytotoxic and non-hemolytic.

Gel and Degradation Time Measurements

The gel time for all cases was measured starting from the addition ofthe ester until the gelation of the solution. The gel point was noted bypipetting 1 mL of the reaction mixture and observing the dropwiseincrease in viscosity until the mixture ceased to flow. Degradation ofthe polymers was performed by the addition of 1 to 10 mL of phosphatebuffered saline per 1 g of the material in a 50 mL centrifuge tube andincubating the mixture at 37° C. A digital water bath was used tomaintain the temperature. The degradation time was measured startingfrom the day of addition of the phosphate buffer to complete dissolutionof the polymer into solution.

The effect of reaction buffer pH, phosphate concentration, polymerconcentration and reaction temperature on the gel times werecharacterized. The buffer pH was varied from 7.2 to 8.0 by the dropwiseaddition of either 50% aqueous NaOH or 6.0 N HCl. Phosphateconcentrations of 0.01, 0.02 and 0.05 M were prepared and adjusted to pH7.4. Polymer concentrations from 2 to 20% solution were studied.Reaction temperatures of 5, 20, and 37° C. were tested by keeping themonomers, buffers, and reaction mixture at the appropriate temperature.The 5° C. environment was provided by a refrigerator and the 37° C.temperature was maintained via the water bath. Room temperature wasfound to be 20° C.

The effect of degradation buffer pH and the proportion of degradableamine in the polymer formulation on the degradation times were explored.The degradation buffer pH was varied from 7.2 to 9.0 by the dropwiseaddition of either 50% aqueous NaOH or 6.0 N HCl. The degradable aminecomponents studied were either the 4ARM-20k-AA or the 8ARM-20k-AA, andthe percent of degradable amine relative to the non-degradable amine wasvaried from 50 to 100%.

The degradation time is largely dependent on the buffer pH, temperature,and the monomers used. Degradation occurs primarily through ester bondhydrolysis; in biological systems, enzymatic pathways may also play arole. FIG. 8 compares the degradation times of formulations with4ARM-20k-AA and 8ARM-20k-AA in varying amounts. In general, increasingthe amount of degradable acetate amine in relation to the non-degradableamine decreases the degradation times. Additionally, in some instances,the 8ARM-20k-AA exhibits a longer degradation time than the 4ARM-20k-AAper mole equivalent, which becomes especially apparent when the percentof acetate amine drops below 70%.

FIG. 9 shows the effect of the buffer pH on the degradation time. The pHrange between 7.2 and 9.0 was studied. In general, a high pH environmentresults in a greatly accelerated degradation. For example, an increasein pH from approximately 7.4 to 7.7 decreases the degradation time byabout half.

The monomers used in the formulations have also been found to play arole in the way the polymer degrades. For the 8ARM-20k-AA/8ARM-20k-NH2(70/30) & 4ARM-20k-SGA polymer, degradation occurred homogeneouslythroughout the material, resulting in a “smooth” degradation process,which is depicted in FIGS. 10A, 10B, 10C, and 10D. The initial state ofthe polymer is shown in FIG. 10A. The polymer absorbed water and swelledslightly over the initial few days (FIG. 10B). Then, the polymer becamegradually softer yet maintained its shape (FIG. 10C). Finally, thepolymer lost its shape and became a highly viscous fluid (FIG. 10D). The70/30 formulation was chosen for the 14 day ophthalmic applicationdespite its 21 day degradation time because the polymer maintained itsshape up to day 14. From day 14 up to day 21, the polymer began to loseits shape and entered the viscous fluid stage.

Examples of fragmenting degradation processes are shown in FIGS. 11A and11B. When the amount of degradable amine becomes low, non-degradableregions in the polymer may occur. FIG. 11A depicts the8ARM-20k-AA/8ARM-20k-NH2 (60/40) & 4ARM-20k-SGA formulation afterapproximately 80 days. FIG. 11B depicts the 4ARM-20k-AA/8ARM-20k-NH2(70/30) & 4ARM-20k-SGA formulation, which degraded into several largefragments. For applications where the polymers are subjected to greatforces, fragmentation may also occur as the polymer becomes softer andweaker over time.

Polymer Concentration

More dilute polymer solutions may be employed with minimal changes inthe mechanical properties. For the formulation8ARM-20k-AA-20K/8ARM-20k-NH2 (75/25) with 4ARM-20k-SGA and 0.3% HPMC,polymer concentrations of 3.0, 3.5 and 4.0% were studied. FIG. 12A showsthe gel times, which increased steadily as the polymer concentration waslowered. The firmness decreased slightly as the polymer concentrationwas lowered (FIG. 12B). The tack is shown in FIG. 12C. There wasessentially no change in the polymer adhesive properties. The elasticmodulus decreased slightly as the polymer concentration was lowered(FIG. 12D). The swelling or water uptake is shown in FIG. 12E.

TABLE 8 (A) Reaction details for specific sticky formulation; (B)formulation results for a specific sticky formulation with a variety ofviscosity enhancing agents (the hydrogel surface spread test isconducted on a hydrophilic hydrogel surface composed of 97.5% water atan angle of approximately 30°; one drop of the polymer solution from a22 gauge needle is applied to the surface before gelation); (C) theclarity of solutions containing a variety of viscosity enhancing agents,as measured by the % transmission at 650 nm. (A) Components MW wt (g)Arm mmoles Arms Eq % Solution 8ARM-20k-NH2 20000 0.04 8 0.002 0.0164ARM-20k-SGA 20000 0.08 4 0.004 0.016 Phosphate buffer 2.5 mL 0.10M, pH7.80 4.8 (B) Gel Hydrogel Viscous Agent Approximate Time Surface Spread% (w/w) Viscosity (cP) (s) Test Category Notes 0 (Original 1.1 80 2Rigid, has “bounce”. Slight Formulation) elasticity.   5% PVP 1 to 5  902 to 3 No change, except for a slight increase in elasticity.   10% PVP3 to 5  90 2 to 3 Slightly opaque, moderate increase in elasticity.Slippery.   15% PVP 5 to 10 100 2 to 3 Opaque, definite increase inelasticity. Slippery when wet, slightly sticky when dry.   20% PVP 10110 2 Opaque, definite increase in elasticity. Slippery when wet, verysticky when dry.  0.3% HPMC 8.4 80 2 No change.  1.0% HPMC 340.6 90 1 Nochange. 1.25% HPMC 1,000 90 1 No change.  1.5% HPMC 2,000 100 1 Slightlysofter, lacks “bounce”.  2.0% HPMC 4,000 100 1 Slightly softer, lacks“bounce”. Slippery. (C) Sample % Transmission @ 650 nm 0.10M phosphatebuffer, pH 7.80 100.0%  10% PVP  99.9% 1.5% HPMC  95.7% 1.0% HPMC  96.8%0.5% HPMC  99.1% 0.1% HPMC  99.6% Hydrogel Surface Spread TestCategories: 1 No spreading, tight drops that stay in place; 2 Mildspreading, drops drip slowly down; 3 Severe spreading, drops completelywet surface. Water is in category 3.

Methylcellulose (MC) was found to behave similarly to hypromellose(HPMC) and provided workable viscous solutions in the concentrationrange of 0 to 2% (w/w). However, the HPMC dissolved more readily thanthe MC, and the HPMC solutions possessed greater optical clarity; thusthe use of HPMC was favored. Povidone (PVP) dissolved easily in thebuffer, but provided minimal viscosity enhancement even at 20% (w/w).

For the most part, the polymers remain unchanged by the addition of lowconcentrations of HPMC or PVP. However, there was a noticeable change inthe polymer around 0.3% HPMC that was characterized by an enhancedelasticity, as evidenced by the ability of the material to elongate morethan usual without breakage. Above 1.5% HPMC, the polymer becameslightly softer and exhibited less bounce. The gel times also remainedwithin 10 seconds of the gel time for the formulation with no viscousagent. In the case of PVP, significant changes in the polymer occurredabove 10% PVP. The polymer became more opaque with a noticeable increasein elasticity and stickiness. At 15% to 20% PVP, the polymer becamesimilar to the sticky materials, but with a better mechanical strength.The gel times also increased by roughly 20 seconds relative to theformulation with no viscous agent. Thus, the addition of lowerconcentrations of PVP or HPMC to the polymer solutions may be beneficialin improving the polymer's elasticity and lubricity.

The results of the hydrogel surface spread test show that mostformulations belong in category 2.

Based on the these observations, a formulation utilizing 0.3% HPMC waschosen for further evaluation. Above 1.0% HPMC, the solutions becamesignificantly more difficult to mix and dissolution of the monomersbecame an issue. At 0.5% HPMC and above, the formation of air bubblesduring mixing became significant. Furthermore, the solutions were noteasily filtered through a 0.5 μm syringe filter to remove the bubbles.However, the 0.3% HPMC solution was easily filtered even after moderatemixing, resulting in a bubble-free, optically clear polymer.

Viscosity Measurements

The viscosities of the resulting buffer solutions were measured with theappropriately sized Cannon-Fenske viscometer tube from Ace Glass.Viscometer sizes used ranged from 25 to 300. Measurements of selectsolutions were performed in triplicate at both 20° C. and 37° C. Theresults are shown in Table 8B. To calculate the approximate dynamicviscosities, it was assumed that all the buffer solutions had the samedensity as water.

To characterize the rheology of the polymers during the gelationprocess, a size 300 viscometer was used with a formulation that wasdesigned to gel after approximately 15 minutes. The formulation usedinvolved the 8ARM-20k-NH2 with the 4ARM-20k-SGA ester at 2.5% solutionand 0.3% HPMC. The reaction occurred in a 0.05 M phosphate buffer at apH of 7.2. Thus, one viscosity measurement with the size 300 viscometerwas obtained in about one minute and subsequent measurements may beobtained in quick succession up to the gel point.

Hydrogel Surface Spread Test

Since the surface of the retina is extremely hydrophilic, making itlikely that a liquid drop will spread beyond the desired site ofadministration, the spread was modeled using an extremely hydrophilicsurface. To model the performance of the polymer solutions on ahydrophilic surface the extent of spreading and dripping of droplets ona high water content hydrogel surface at an incline of about 30° wasrecorded. The hydrogel was made by dissolving 0.10 g (0.04 mol arm eq.)of 8ARM-20k-NH2 in 7 mL 0.05 M phosphate buffer at pH 7.4 in aPetri-dish, followed by the addition of 0.075 g (0.04 mol arm eq.) of8ARM-15k-SG ester. The solution was stirred with a spatula for 10 to 20seconds and allowed to gel, which typically took 5 to 10 minutes. Thewater content of the resulting polymer was 97.5%.

The test was performed by first preparing the polymer solution in theusual fashion. After thorough mixing, the polymer solution was dispenseddropwise through a 22 gauge needle onto the hydrogel surface. Theresults are shown in Table 8B and were divided into three generalcategories: 1) no spreading, tight drops that stay in place; 2) mildspreading, drops drip slowly down; 3) severe spreading, drops completelywet surface. Water is in category 3.

Swelling Measurements

The extent of swelling in the polymers during the degradation processwas quantified as the liquid uptake of the polymers. A known mass of thepolymer was placed in PBS at 37° C. At specified time intervals, thepolymer was isolated from the buffer solution, patted dry with papertowels and weighed. The percent increase in the mass was calculated fromthe initial mass.

The percent of water uptake by the 8ARM-20k-NH2/4ARM-20k-SGA polymerswith 0, 0.3 and 1.0% HPMC is shown in FIG. 13. The 1.0% HPMC polymerabsorbed up to 30% of its weight in water until day 20. After day 20,the polymer returned to about 10% of its weight in water. In comparison,the 0% HPMC polymer initially absorbed up to 10% of its weight in water,but began to lose water gradually, hovering about 5% of its weight inwater. The 0.3% HPMC polymer behaved in an intermediate fashion. Itinitially absorbed up to 20% of its weight in water, but returned toabout 10% of its weight in water after a week and continued to slowlylose water.

Specific Gravity Measurements

The specific gravity of the polymers was obtained by preparing thepolymer solution in the usual fashion and pipetting 1.00 mL of thethoroughly mixed solution onto an analytical balance. The measurementswere performed in triplicate at 20° C. The specific gravity wascalculated by using the density of water at 4° C. as the reference.

The obtained values for the specific gravity are shown in FIG. 14. Thespecific gravity of the polymers did not differ significantly from thatof the buffer solution only, both of which were essentially the same asthe specific gravity of water. Exceptions may occur when the polymersolution is not filtered and air bubbles become embedded in the polymermatrix.

Barium Sulfate Suspensions

For imaging purposes, barium sulfate was added to several polymerformulations as a radiocontrast agent. Barium sulfate concentrations of1.0, 2.0, 5.0 and 10.0% (w/v) were explored. The viscosity of theresulting polymer solutions was measured and the effect of bariumsulfate addition on the polymer gel times and syringabilitycharacteristics were also studied.

Barium sulfate concentrations of 1.0, 2.0, 5.0 and 10.0% (w/v) wereexplored. The opaque, milky white suspensions formed similarly opaqueand white polymers. No changes in the gel times were observed.Qualitatively, the polymers appeared to have similar properties to thatof polymers without barium sulfate. All formulations were able to bereadily dispensed through a 22 gauge needle.

The results of the viscosity measurements for barium sulfateconcentrations of 1.0, 2.0, 5.0 and 10.0% are shown in FIG. 15. Theviscosity remained relatively stable up to 2.0%; at 5.0%, the viscosityincreased slightly to about 2.5 cP. There was a sharp increase in theviscosity to nearly 10 cP as the concentration approached 10.0%. Thus, abarium sulfate concentration of 5.0% was chosen as a balance betweenhigh contrast strength and similarity to unmodified polymerformulations.

Hydrogel Firmness, Elastic Modulus, and Adhesion

The firmness of the polymers was characterized by a Texture Analyzermodel TA.XT.plus with Exponent software version 6.0.6.0. The methodfollowed the industry standard “Bloom Test” for measuring the firmnessof gelatins. In this test, the TA-8¼″ ball probe was used to penetratethe polymer sample to a defined depth and then return out of the sampleto the original position. The peak force measured is defined as the“firmness” of the sample. For the polymers studied, a test speed of 0.50mm/sec, a penetration depth of 4 mm, and a trigger force of 5.0 g wereused. The polymers were prepared on a 2.5 mL scale directly in a 5 mLsize vial to ensure consistent sample dimensions. The vials used wereThermoScientific/Nalgene LDPE sample vials, product#6250-0005(LOT#7163281060). Measurements were conducted at 20° C. The polymerswere allowed to rest at room temperature for approximately 1 hour beforemeasuring. Measurements were performed in triplicate for at least threesamples. A sample plot generated by the Exponent software running thefirmness test is given in FIG. 16. The peak of the plot represents thepoint at which the target penetration depth of 4 mm was reached.

The elastic modulus of the polymers was characterized by a TextureAnalyzer model TA.XT.plus with Exponent software version 6.0.6.0. Inthis test, the TA-19 Kobe probe was used to compress a polymer cylinderof known dimensions until fracture of the polymer occurs. The probe hasa defined surface area of 1 cm². The modulus was calculated as theinitial slope up to 10% of the maximum compression stress. For thepolymers studied, a test speed of 5.0 mm/min and a trigger force of 5.0g were used. The sample height was auto-detected by the probe. Thepolymers were prepared on a 2.5 mL scale directly in a 5 mL size vialcap to ensure consistent sample dimensions. The vials used wereThermoScientific/Nalgene LDPE sample vials, product#6250-0005(LOT#7163281060). Measurements were conducted at 20° C. The polymerswere allowed to rest at room temperature for approximately 1 hour beforemeasuring. Measurements were performed for at least three samples. Asample plot generated by the Exponent software running the modulus testis given in FIG. 17. The polymers typically behaved elastically for theinitial compression, as evidenced by the nearly linear plot.

The adhesive properties of the polymers were characterized by a TextureAnalyzer model TA.XT.plus with Exponent software version 6.0.6.0. In theadhesive test, the TA-57R 7 mm diameter punch probe was used to contactthe polymer sample with a defined force for a certain amount of time,and then return out of the sample to the original position. An exemplaryplot generated by the Exponent software running the adhesive test isgiven in FIG. 18. The plot begins when the probe hits the surface of thepolymer. The target force is applied on the sample for a defined unit oftime, represented by the constant force region in the plot. Then, theprobe returns out of the sample to the original position and theadhesive force between the probe and the sample is measured as the“tack”, which is the peak force required to remove the probe from thesample. Other properties that were measured include the adhesion energyor the work of adhesion, and the material's “stringiness.” The adhesionenergy is simply the area under the curve representing the tack force.Thus, a sample with a high tack and low adhesion energy willqualitatively feel very sticky, but may be cleanly removed with a quickpull; a sample with a high tack and high adhesion energy will also feelvery sticky, but the removal of the material will be more difficult andmay be accompanied by stretching of the polymer, fibril formation andadhesive residues. The elasticity of the polymer is proportional to themeasured “stringiness”, which is the distance the polymer stretcheswhile adhered to the probe before failure of the adhesive bond. For thepolymers studied, a test speed of 0.50 mm/sec, a trigger force of 2.0 g,and a contact force of 100.0 g and contact time of 10.0 sec were used.The polymers were prepared on a 1.0 to 2.5 mL scale directly in a 5 mLsize vial to ensure consistent sample surfaces. The vials used wereThermoScientific/Nalgene LDPE sample vials. Measurements were conductedat 20° C. The polymers were allowed to rest at room temperature forapproximately 1 hour before measuring. As reference materials, theadhesive properties of a standard Post-It Note® and Scotch Tape® weremeasured. All measurements were performed in triplicate. The averagesand standard deviations were calculated.

The effect of HPMC addition to the mechanical properties of the polymerswas explored, along with the effect of adding degradable 8ARM-20k-AAamine. The results are shown in FIGS. 19A, 19B, 20A, and 20B. Under thestated conditions of the firmness test, it was found that the additionof 0.3% HPMC decreased the firmness of the polymer by about half (FIG.19A). This corresponds to a slight decrease in the elastic modulus (FIG.20A). The 1.0% HPMC polymer had approximately the same firmness as the0.3% HPMC polymer, but a slight decrease in the elastic modulus. Thedisparity between the firmness and modulus tests is likely due toexperimental error. The polymer solutions were not filtered, so thepresence of air bubbles likely increased the errors. The water contentof the polymers may also change as the polymers were sitting in the air,essentially changing the physical properties of the materials.

It was found that the addition of the degradable 8ARM-20k-AA amine didnot substantially change the measured values of the firmness or theelastic modulus (FIG. 19B and FIG. 20B). The results of the adhesiontesting are shown in FIGS. 21A, 21B, 21C, and 21D. The measured valuesfor a standard commercial Post-It™ Note are also included as areference. The polymer tack was found to be around 40 mN, which is aboutthree times less than that of a Post-It™ Note. The adhesive propertiesof the polymer were not found to vary with the addition of thedegradable amine.

FIG. 22 shows the firmness vs. degradation time for the8ARM-20k-AA/8ARM-20k-NH2 (70/30) & 4ARM-20k-SGA at 4.8% solution with0.3% HPMC. The error bars represent the standard deviations of 3samples. The degradation time for the polymer was 18 days. The firmnessof the polymer strongly correlated with the extent of degradation.Swelling may also play a role during the early stages.

Optical Clarify

A Thermo Scientific GENESYS 10S UV-Vis spectrophotometer was used tomeasure the optical clarity of the viscous solutions. To a quartzcuvette, 1.5 mL of the sample solution was pipetted. The buffer solutionwith no additives was used as the reference. The stable % transmissionof the sample was recorded at 650 nm and the results are listed in Table8C.

All of the viscous solutions under consideration were found to haveacceptable to excellent optical clarity under the concentration rangesused (greater than 97% transmission). For the highly viscous solutions,air bubble formation during mixing was observed, which may be resolvedby the addition of an anti-foaming agent, or through the use of asyringe filter.

Example 12 General Procedure for the Preparation of In VivoPolymerizable Thin Films

Several representative formulations for both sticky and non-sticky filmsare listed in Table 9 along with specific reaction details. The filmshad thicknesses ranging from 100 to 500 μm, and may be layered withdifferent formulations in a composite film.

TABLE 9 (A) Summary of the reaction details for several representativethin film formulations; (B) more detailed tabulation of a selection ofthe reaction details including moles (films ranged in thickness from 100to 500 μm). (A) Amine/Ester Components Molar Ratio Buffer % Solution4ARM-20k-AA & 8ARM-15k-SG 1 0.15M phosphate, 19.6 pH 7.99 4ARM-5k-NH2 &4ARM-10k-SG 4.5/1   0.05M phosphate, 39 pH 7.40 4ARM-5k-NH2 &4ARM-10k-SG 1 0.05M phosphate, 36.4 pH 7.40 4ARM-5k-NH2 & 4ARM-10k-SG &4.5/1   0.10M phosphate, 39 HPMC (1.25%) pH 7.80 4ARM-2k-NH2 &4ARM-10k-SG & 8/1 0.10M phosphate, 30.6 HPMC (1.5%) pH 7.80 4ARM-2k-NH2& 4ARM-20k-SGA & 8/1 0.15M phosphate, 30 MC (2%) pH 7.94 4ARM-2k-NH2 &4ARM-20k-SGA & 10/1 0.15M phosphate, 30 MC (2%) pH 7.94 (B) Polymer % WtArms Solution Components MW Mmoles (g) Arm mmoles Eq (w/v) 4ARM-20k-AA20000 1000 0.2 4 0.01 0.04 8ARM-15k-SG 15000 1000 0.075 8 0.01 0.04Buffer Volume (phosphate) 1.4 19.6 4ARM-5k-NH2 5000 1000 0.27 4 0.050.22 4ARM-10k-SG 10000 1000 0.12 4 0.01 0.05 Buffer Volume (phosphate) 139.0 4ARM-5k-NH2 5000 1000 0.17 4 0.03 0.14 4ARM-10k-SG 10000 1000 0.344 0.03 0.14 Buffer Volume (phosphate) 1.4 36.4 4ARM-5k-NH2 5000 10000.27 4 0.05 0.22 4ARM-10k-SG 10000 1000 0.12 4 0.01 0.05 Buffer Volume(phosphate) 1 39.0 Viscosity Enhancer 1.25% HPMC

Example 13 Preparation of Kits and their Use

Several kits were prepared with polymer formulation tested earlier. Thematerials used to assemble the kits are listed in Table 10 and theformulations used are listed in Table 11. The kits are typicallycomposed of two syringes, one syringe containing the solid componentsand the other syringe containing the liquid buffer. The syringes areconnected via a mixing tube and a one-way valve. The contents of thesyringes are mixed via opening the valve and transferring the contentsof one syringe into the other, repeatedly, for 10 to 20 seconds. Thespent syringe and mixing tube are then removed and discarded, and theactive syringe is fitted with a dispensing unit, such as a needle orcannula, and the polymer solution is expelled until the onset ofgelation. In other embodiments, the viscous solution impedes thedissolution of the solid components and thus a third syringe isemployed. The third syringe contains a concentrated viscous buffer thatenhances the viscosity of the solution once all the components havedissolved. In some embodiments, the optical clarity of the resultingpolymer is improved through the addition of a syringe filter.

All of the formulations tested were easily dispensed through a 22 gaugeneedle. The mixing action between the two syringes was turbulent and theintroduction of a significant amount of air bubbles was apparent. Gentlemixing results in a clear material free of bubbles. Alternatively, theuse of a syringe filter was found to remove bubbles without any changein the polymer properties.

TABLE 10 Materials used to fabricate kits including vendor, part numberand lot number. Description Vendor Vincon Tubing, ⅛″ I.D. ¼″ O.D. RyanHerco Flow 1/16″ wall, 100 Ft. Solutions 12 mL Leur-Lok Syringe TycoHealthcare, Kendall Monoject ™ 3 mL Leur-Lok Syringe Tyco Healthcare,Kendall Monoject ™ One Way Stopcock, Female Luer Lock to Male LuerQOSINA Female Leur Lock Barb for ⅛″ I.D. tubing, RSPC QOSINA Non-ventedLuer Dispensor Tip Cap, White QOSINA 32 mm Hydrophilic Syringe Filter, 5micron PALL ®Life Sciences

TABLE 11 The detailed contents for four different kits; the solidcomponents are in one syringe, while the liquid components are inanother syringe; a mixing tube connects the two syringes. Components MWwt (g) Arm mmoles Arms Eq % Solution 8ARM-20k-NH2 20000 0.04 8 0.0020.016 4ARM-20k-SGA 20000 0.08 4 0.004 0.016 Phosphate buffer 2.5 mL0.10M, pH 7.80 4.8 Viscosity Enhancer No viscosity enhancer 8ARM-20k-NH220000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 0.08 4 0.004 0.016 Phosphatebuffer 2.5 mL 0.10M, pH 7.80 4.8 Viscosity Enhancer 0.3% HPMC8ARM-20k-NH2 20000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 0.08 4 0.0040.016 Phosphate buffer 2.5 mL 0.10M, pH 7.80 4.8 Viscosity Enhancer 7.5%Povidone 8ARM-20k-NH2 20000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 0.08 40.004 0.016 Phosphate buffer 2.5 mL 0.10M, pH 7.80 4.8 ViscosityEnhancer 1.0% HPMC

Several additional kits were prepared with the polymer formulation thatperformed the best in initial trials. The materials used to assemble thekits are listed in Table 12. The kits are typically composed of twosyringes, one syringe containing the solid components and the othersyringe containing the liquid buffer. The syringes were loaded byremoving the plungers, adding the components, purging the syringe with agentle flow of nitrogen gas for 20 seconds, and then replacing theplunger. Finally, the plungers were depressed as much as possible toreduce the internal volume of the syringes. The specifications for theamounts of chemical components in the kits are listed in Table 13A. Asummary describing the lots of kits prepared is listed in Table 13B.

The syringes were connected directly after uncapping, the male partlocking into the female part (FIGS. 23A and 23B). The contents of thesyringes were mixed via transferring the contents of one syringe intothe other, repeatedly, for 10 to 20 seconds. The spent syringe was thenremoved and discarded, and the active syringe was fitted with adispensing unit, such as a needle or cannula, and the polymer solutionwas expelled until the onset of gelation. In other embodiments, theviscous solution impeded the dissolution of the solid components andthus a third syringe was employed. The third syringe contained aconcentrated viscous buffer that enhanced the viscosity of the solutiononce all the components had dissolved.

All the formulations tested were easily dispensed through a 22 gaugeneedle. The mixing action between the two syringes was turbulent and theintroduction of a significant amount of air bubbles was apparent. Theuse of a syringe filter was found to remove bubbles without any changein the polymer properties.

The prepared kits were placed into foil pouches along with one oxygenabsorbing packet per pouch. The pouches were heat sealed with a CHTC-280PROMAX tabletop chamber sealing unit. Two different modes of sealingwere explored: under nitrogen and under vacuum. The settings for sealingunder nitrogen were: 30 seconds of vacuum, 20 seconds of nitrogen, 1.5seconds of heat sealing, and 3.0 seconds of cooling. The settings forsealing under vacuum were: 60 seconds of vacuum, 0 seconds of nitrogen,1.5 seconds of heat sealing, and 3.0 seconds of cooling.

TABLE 12 Materials used to fabricate kits including vendor, part numberand lot number. Description Vendor 12 mL Male Luer-Lok Syringe TycoHealthcare, Kendall Monoject ™ 5 mL Female Luer Lock Syringe, PurpleQOSINA Male Luer Lock Cap, Non-vented QOSINA Female Non-vented LuerDispensor Tip Cap, QOSINA White 100 cc oxygen absorbing packet IMPAK6.25″ × 9″ OD PAKVF4 Mylar IMPAK foil pouch

TABLE 13 Specifications for kit components for the 8ARM-20k-AA/8ARM-20-NH2 & 4ARM-20k-SGA formulation with 60, 65, 70 and 75% degradableamine (A). LOT formulation summary (B). (A) Specifications Components60/40 65/35 70/30 75/25 8ARM-20k-AA 0.024-0.026 g 0.026-0.027 g0.028-0.029 g 0.030-0.031 g 8ARM-20k-NH2 0.014-0.016 g 0.013-0.014 g0.011-0.012 g 0.009-0.010 g 4ARM-20k-SGA 0.080-0.082 g 0.080-0.082 g0.080-0.082 g 0.080-0.082 g Phosphate 2.50 mL of 0.10M phosphate, pH7.58, 0.30% HPMC Buffer (8.48 cSt +/− 0.06 @ 20° C.) (B) FormulationBuffer pH Sealing Method Notes 60/40 7.46 nitrogen 60/40 7.58 nitrogen60/40 7.72 nitrogen 70/30 7.58 vacuum 70/30 7.58 vacuum no nitrogenpurging of syringe 65/35 7.58 vacuum 75/25 7.58 vacuum 75/25 7.58 vacuum75/25 7.58 nitrogen 65/35 7.58 vacuum 65/35 7.58 nitrogen

Example 14 Retinal Patch in Harvested Pig Eyes

The formulations listed in Table 14 were used and tested to assesspolymers with respect to adherence, stickiness, thickness, andtransparency in harvested pig eyes. A 0.15 M phosphate buffer was madeby dissolving 9.00 g (0.075 mol) NaH₂PO₄ in 500 mL of distilled water at25° C. with magnetic stirring. The pH was then adjusted to 7.99 with thedropwise addition of 50% aqueous NaOH. Phosphate buffered saline (PBS)was prepared by dissolving two PBS tablets (Sigma Chemical, P4417) in400 mL of distilled water at 25° C. with vigorous shaking. The solutionhas the following composition and pH: 0.01 M phosphate, 0.0027 Mpotassium chloride, 0.137 M sodium chloride, pH 7.46.

TABLE 14 Components of Formulations Tested in Harvested Pig Eyes. Arms %Formulation Components MW wt (g) Arm mmoles Eq Solution A 4ARM-5k-NH25000 0.2 4 0.04 0.16 4ARM-10k-SG 10000 0.08 4 0.008 0.032 Phosphatebuffer 1.5 mL 0.15M, 18.7 pH 7.99 B 4ARM-5k-NH2 5000 0.3 4 0.06 0.244ARM-10k-SG 10000 0.12 4 0.012 0.048 Phosphate buffer 1.5 mL 0.10M, 28pH 7.80 C 8ARM-20k-NH2 20000 0.04 8 0.002 0.016 4ARM-20k-SGA 20000 0.084 0.004 0.016 Phosphate buffer 2.5 mL 0.10M, 4.8 pH 7.80

The general design and of the delivery device is shown in FIG. 24. Thekits are composed of two syringes, one syringe containing the solidcomponents and the other syringe containing the liquids. The syringesare connected via a mixing tube and a one-way valve. The contents of thesyringe are mixed after opening the valve and transferring the contentsof one syringe into the other, repeatedly, for 10 to 20 seconds. Thespent syringe and mixing tube are then removed and discarded, and theactive syringe is fitted with a needle, and the polymer solution isinjected into the desired site while still in the liquid state. Theliquid polymer turns in to a solid at a pre-set time at the desired siteand sticks to the tissue. The optical clarity of the resulting polymeris improved by removing the air bubbles through the addition of asyringe filter.

Retinal Tissue Tests

Pig eyes were obtained and stored appropriately. Surgery was carried outcarefully and the vitreous humor was separated from the retina and thenthe retina was cut at several places and then pinned appropriately to aStyrofoam base to create basically a flat retinal surface. The liquidsuture formulations of Table 14 were mixed per the mixing procedureabove and carefully deposited drop by drop over the retinal surface. Anyspreading of the drop from the location was carefully observed andrecorded. After test samples gelled in about 60-120 seconds, the bondstrength of the adhesion was evaluated. All three formulations wereapplied the same way. The results are summarized in Table 15.

TABLE 15 Test Results for Formulations A-C on Retinal Tissue.Formulation Results/Observations A It is a sticky material, gelled in110 seconds. Not deemed “hard” enough. The material showed bubbles andit spread all over; not localized B It is a sticky material, gelled in120 seconds. Not deemed “hard” enough. The material showed bubbles andit spread all over; not localized. C It is a less sticky material,gelled in 80 seconds. Bonding was satisfactory. Formulation bonded toretina uniformly and showed no delamination area under the microscope.The material showed bubbles and was also not localized only at thetarget site. Bubbles in the polymer lowered the optical clarity at thelocation. It also passed the blue dye leak test indicating that thebonding was strong and leak free.

Of the 3 formulation types tested, formulation “C” was most successfuleven though all 3 formulations provide satisfactory results.

In order to improve the clarity and control the polymer spread, bubbleformation and drop spread were evaluated.

In order to avoid bubble formation, the mixing procedure was altered,wider diameter mixing tubes were used, the syringe size was changed,antifoaming agents were added, organic solvents such as DMSO were addedto the formulation, or filters were used. After examining all options,the use of 5 microns filter was deemed to be the most effective andpractical procedure. The bubble formation was eliminated by using a 5micron filter during the injection process. 0.2 Micron filter also wasacceptable except for higher viscosity materials which clogged thefilter.

In some instances, a higher viscosity material does not spread as fastas the lower viscosity material. Therefore, several viscosity enhancingagents were formulated with the initial formulations as shown in Table16. The initial results for viscosity are included in Table 11. Opticalclarity data is included in Table 8.

TABLE 16 Viscosity Enhanced Formulations A-C Arms % FormulationComponents MW wt. (g) Arm mmoles Eq Solution C-1 8ARM-20k-NH2 20000 0.048 0.002 0.016 4ARM-20k-SGA 20000 0.08 4 0.004 0.016 Phosphate buffer 2.5mL YC-06-105 (0.10M, pH 7.80) 4.8 Viscosity Enhancer No viscosityenhancer C-2 8ARM-20k-NH2 20000 0.04 8 0.002 0.016 4ARM-20k-SGA 200000.08 4 0.004 0.016 Phosphate buffer 2.5 mL YC-06-105 (0.10M, pH 7.80)4.8 Viscosity Enhancer 0.3% HPMC C-3 8ARM-20k-NH2 20000 0.04 8 0.0020.016 4ARM-20k-SGA 20000 0.08 4 0.004 0.016 Phosphate buffer 2.5 mLYC-06-105 (0.10M, pH 7.80) 4.8 Viscosity Enhancer 1.0% HPMC

20 pig eyes were used in the study. After a 3-port pars plana vitrectomy(after cataract extraction by phaco) using a 23 gauge transconjunctivaltechnique, a retinal tear or hole was created. The in vivo gellingophthalmic pre-formulation was delivered through a 23 gauge intraocularcannula after an air/fluid exchange to create a retinal patch over theretinal lesion. It was verified that the polymer covered the retinaltear area and created a patch on the damaged retina. After about 1minute the liquid became sold and adhered to the pathologic area of theretina. A stained liquid (trypan blue) was injected in the sub-retinalspace to assess the resistance of the patch. The optical clarity metexpectation.

Example 15 Retinal Patch in Live Animals (Rabbits)

This study is designed to select the best polymer in terms of adherence,stickiness, thickness, and transparency; to improve the design of thedelivery system and technique; assess the efficacy of the retinal patchin keeping the retinal patch attached; and to assess the safety of thepolymer in terms of inflammatory reactions or other toxic effects on theretina.

40 Rabbits are used in the study, wherein 40 eyes are subject totreatment while the fellow eye is used as the control eye. After a3-port pars plana vitrectomy (after cataract extraction by phaco), aretain tear or hole is created. The in vivo gelling ophthalmicpre-formulation is delivered to create a retinal patch over the retinallesion in 20 rabbits, while the entire vitreous cavity is filled withthe polymer in the other 20 rabbits.

The eyes are evaluated for intraocular inflammation at day 1, 7, 15, and30 by slit lamp evaluation, fundus evaluation, and intraocular pressuremeasurements. The eyes are evaluated for retinal toxicity at day 1, 7,15, and 30 by anatomopathologic evaluation of the retina (which has orhas not been in contact with the patch), multifocal ERG and +/−DNAanalysis.

Example 16 Method of Treating Retinal Tear Using Local Anesthesia

This study is designed to treat a retinal tear using local anesthesia.The site of the hole, tear, or retinal detachment is identified usingthe existing and well established techniques. Once thehole/tear/detachment location is identified, the kits are preparedaccording to Example 13. Using a sharp 24 to 28 gauge needle, about10-500 micro liters of the polymers is injected around and over the holemaking sure that the entire surface area is fully covered with thepolymer mixture. The viscosity of the reacting mixture does not allowexcessive flow of the fluid past the site of the injection. Everyattempt is made to keep the polymer injection near the hole, tear, ordetachment sites by keeping the subject in a stable position. Even if asmall amount flows over in another area, it will dissolve and disappearin about 14-17 days. After the tear is covered, the polymer solidifiesin less than 3 minutes from the time the mixing was started. The polymerwill stay at the site for 14-17 days and then dissolve and disappear.

Example 17 Clinical Trial for the Treatment of Retinal Detachment with aHydrogel Formulation after Surgery

The aim of the study is to evaluate the influence of exemplary hydrogelformulation C on thickness of the retinal nerve fiber layer by usingoptical coherence tomography (OCT) in patients following pars planavitrectomy. The study will include 60 patients with a Formulation Cprovided herein who will be surgically treated with pars planavitrectomy for rhegmatogenous retinal detachment and proliferativevitreoretinopathy. All subjects will be subdued to completeophthalmologic examinations, measurements of the retinal nerve fiberlayer thickness by an OCT examination, tests of the visual field withthe use of an Octopus computed perimeter (automated static perimetry)and FDT-perimetry—both prior to the surgery, and on control visits forcheck-up during the postoperative period. All results provided bypostoperative examinations will be compared with one another. The studyis expected to provide data on the effect of an exemplary Formulation Con thickness of the retinal nerve fiber layer. It is also planned toshow possibilities and advantages of OCT as a method of choice in thefollow-up of patients with an intraocular Formulation C.

Condition Intervention Rhegmatogenous Retinal Other: Optical coherencetomography Detachment Side Effect Drug: Local medical treatment of ofFormulation C raised intraocular pressure

Study Type: Observational. Study Design: Observational Model: CaseControl.

Time Perspective: Prospective.

Eligibility

Ages Eligible for Study: 18 Years to 80 Years

Genders Eligible for Study: Both

Accepts Healthy Volunteers: No

Sampling Method: Probability Sample Criteria

Inclusion Criteria:—patients with rhegmatogenous retinal detachment

Exclusion Criteria:

-   -   preexistent glaucoma    -   previous retinal surgery    -   placement of scleral buckle during surgery        Further Study Details:

Primary Outcome Measures: Evidence of Retinal Nerve Fibre LayerThickness Change Measured by Optical Coherence Tomography, Time Frame: 6months.

Retinal nerve fiber layer thickness change measured by optical coherencetomography might be an additional parameter that could provide newinsights into clinical decision making in patients with exemplaryformulation C.

Secondary Outcome Measures: Retinal Nerve Fiber Layer Thickness Changein Patients With Raised Intraocular Pressure Secondary to Formulation C,Time Frame: 6 months.

To assess whether retinal nerve fiber layer thickness changes inpatients with raised intraocular pressure secondary to Formulation C.

Groups/Cohorts Assigned Interventions Patients without Optical coherencetomography will be performed in raised IOP all study patients followingpars plans vitrectomy and Formulation C. A fellow eye of each patientwill serve as a control. Each patient enrolled in a study will receive 4measurements: On 7th postoperative day On 30th postoperative day On 90thpostoperative day On 180th postoperative day Patients with Opticalcoherence tomography will be performed in raised IOP all study patientsfollowing pars plans vitrectomy and Formulation C. A fellow eye of eachpatient will serve as a control. Each patient enrolled in a study willreceive 4 measurements: On 7th postoperative day On 30th postoperativeday On 90th postoperative day On 180th postoperative day

Example 18 Clinical Trial of Optical Coherence Tomography with aFormulation C Filled Eye

The aim of this study is to determine the condition to detect the statusof a macular hole by spectral domain optical coherence tomography(SD-OCT) in Formulation C filled eyes. The macular area is scanned bySD-OCT (OCT-4000, Carl Zeiss Meditec) in the patients who underwentvitreous surgery for macular hole to detect macular hole closure onpostoperative days 1, 3, 7, and 30.

26 eyes are studies with an idiopathic macular hole (MH), 7 eyes with aMH retinal detachment (MHRD), and 4 eyes with a MH with myopic tractionmaculopathy. This is a prospective study. The age, gender, laterality ofthe diseased eye, stage of MH based on the Gass classification, andSnellen best-corrected visual acuity (BCVA) are recorded. The axiallength is measured preoperatively in eyes with MH and postoperatively ineyes with MHRD and myopic traction maculopathy to avoid the effect ofretinal detachment on the axial length. The presence of a posteriorstaphyloma within the posterior vascular arcade is determined byophthalmoscopy and ultrasonography.

Condition Intervention Macular Hole Procedure: Vitreous surgery

Study Type: Observational

Study Design: Observational Model: Cohort

Time Perspective: Prospective

Eligibility

Ages Eligible for Study: 35 Years to 85 Years. Genders Eligible forStudy: Both. Accepts Healthy Volunteers: No. Sampling Method:Non-Probability Sample.

Study Population. The patients who have a vitreous surgery for macularhole, macular hole retinal detachment, myopic traction maculopathy andexamined spectral domain optical coherence tomography preoperatively andpostoperatively.

Criteria

Inclusion Criteria: the patients who had a vitreous surgery for macularhole, macular hole retinal detachment, macular hole with myopic tractionmaculopathy and examined spectral domain optical coherence tomographypreoperatively and postoperatively.

Exclusion Criteria: the patients who had vitreous surgery for otherdisease; the patients who did not have postoperative examination ofspectral domain optical coherence tomography.

Further Study Details

Primary Outcome Measures: macular hole closure detected by spectraldomain optical coherence tomography, Time Frame: Change frompreoperative status up to postoperative day 30. The macular hole closurein eyes with Formulation C is detected by spectral domain opticalcoherence tomography.

Secondary Outcome Measures: preoperative and postoperative vision. Thevision is measured preoperatively and postoperative day 30. Thepreoperative and postoperarive vision are measured.

Groups/Cohorts Assigned Interventions Macular hole Procedure: Vitreoussurgery The patients of idiopathic macular Vitreous surgery is performedto treat hole enrolled in the study the original disease not for thestudy. Macular hole retinal detachment Procedure: Vitreous surgery Thepatients of macular hole Vitreous surgery is performed to treat retinaldetachment enrolled in the original disease not for the study. the studyMyopic traction maculopathy Procedure: Vitreous surgery The patients ofmacular hole with Vitreous surgery is performed to treat myopic tractionmaculopathy the original disease not for the study. enrolled in thestudy

Standard pars plana vitrectomy is performed. The internal limitingmembrane (ILM) is removed after making it visible with triamcinoloneacetonide or indocyanine green in all eyes. Preoperative cataracts aregraded as mild (nuclear sclerosis 1+) or moderate to advanced (nuclearsclerosis 2+ or 3+), and phacoemulsification with implantation of anintraocular lens is performed on all cataractous eyes higher thangrade 1. A Formulation C is used to fill the retina.

All surgery is performed under retrobulbar anesthesia, and a writteninformed consent is obtained from all patients after a full explanationof the purpose and possible complications of the treatment. The entiremacular area is scanned by SD-OCT in the sitting position to avoidmissing a MH. The 5-line raster mode is used to obtain high qualityimages on postoperative days 1, 3, 7, and 30. The ability to detect aclosed MH or the status of the foveal detachment or schisis by theSD-OCT is evaluated, and the pre- and postoperative factors thataffected the OCT images are investigated.

What is claimed is:
 1. An in vivo gelling pre-formulation for the treatment of retinal detachment, comprising: (a) multi-ARM nucleophilic polyol monomers having more than two nucleophilic arms, wherein each nucleophilic arm comprises a polyethyleneglycol chain and terminates in a nucleophilic amino group; wherein the nucleophilic arms of the multi-ARM nucleophilic polyol monomers are selected from

and wherein n is 1-200; (b) multi-ARM electrophilic polyol monomers having more than two electrophilic arms, wherein each electrophilic arm comprises a polyethyleneglycol chain and terminates in an electrophilic succinimidyl group; wherein the electrophilic arms of the multi-ARM electrophilic polyol monomers are selected from

wherein m is 2 or 3; and wherein n is 1-200; and (c) a viscosity enhancer selected from carboxymethylcellulose sodium, ethylcellulose, sodium alginate and mixtures thereof; wherein the viscosity of the in vivo gelling pre-formulation is between about 5 cP and 4000 cP; and wherein the in vivo gelling pre-formulation polymerizes and/or gels at a target site of an eye to form a biocompatible retinal patch.
 2. The in vivo gelling pre-formulation of claim 1, wherein the pre-formulation further comprises a buffer providing a pH range of about 6.0 to about 8.5.
 3. The in vivo gelling pre-formulation of claim 1, wherein the pre-formulation further comprises a therapeutic agent.
 4. The in vivo gelling pre-formulation of claim 1, wherein the multi-ARM nucleophilic polyol monomers are selected from

wherein R is hexaglycerol or tripentaerythritol; and wherein n is 1-200.
 5. The in vivo gelling pre-formulation of claim 1, wherein the multi-ARM electrophilic polyol monomers are selected from

wherein R is hexaglycerol or tripentaerythritol; and wherein n is 1-200.
 6. The in vivo gelling pre-formulation of claim 1, wherein the pre-formulation is prepared from the following multi-ARM polyol monomers:

wherein R is hexaglycerol or tripentaerythritol; and wherein n is such that the molecular weight of each of the polyol monomer is 20 kDa.
 7. A method of treating retinal detachment, comprising delivering an in vivo gelling pre-formulation to a site of a retinal tear in a human eye, the in vivo gelling pre-formulation comprising: (a) multi-ARM nucleophilic polyol monomers having more than two nucleophilic arms, wherein each nucleophilic arm comprises a polyethyleneglycol chain and terminates in a nucleophilic amino group; wherein the nucleophilic arms of the multi-ARM nucleophilic polyol monomers are selected from

and wherein n is 1-200; (b) multi-ARM electrophilic polyol monomers having more than two electrophilic arms, wherein each electrophilic arm comprises a polyethyleneglycol chain and terminates in an electrophilic succinimidyl group; wherein the electrophilic arms of the multi-ARM electrophilic polyol monomers are selected from

wherein m is 2 or 3; and wherein n is 1-200; and (c) a viscosity enhancer selected from carboxymethylcellulose sodium, ethylcellulose, sodium alginate, and mixtures thereof; wherein the viscosity of the in vivo gelling pre-formulation is between about 5 cP and 4000 cP; and wherein the in vivo gelling pre-formulation polymerizes and/or gels at a target site of an eye to form a biocompatible retinal patch.
 8. The method of claim 7, wherein the pre-formulation further comprises a buffer providing a pH range of about 6.0 to about 8.5.
 9. The method of claim 7, wherein the pre-formulation further comprises a therapeutic agent.
 10. The method of claim 7, wherein the multi-ARM nucleophilic polyol monomers are selected from

wherein R is hexaglycerol or tripentaerythritol; and wherein n is 1-200.
 11. The method of claim 7, wherein the pre-formulation is prepared from the following multi-ARM polyol monomers:

wherein R is hexaglycerol or tripentaerythritol; and wherein n is 1 to
 200. 12. The in vivo gelling pre-formulation of claim 1, wherein the viscosity enhancer is ethylcellulose.
 13. The in vivo gelling pre-formulation of claim 1, wherein the viscosity enhancer is selected from sodium alginate, carboxymethylcellulose sodium and mixtures thereof.
 14. The method of claim 7, wherein the viscosity enhancer is ethylcellulose.
 15. The method of claim 7, wherein the viscosity enhancer is selected from sodium alginate, carboxymethylcellulose sodium and mixtures thereof. 